product catalog - eppendorf...scientists around the world are working to leap ahead of diseases and...

biotechrabbit

Product Catalog2015-2016

2 www.biotechrabbit.com

biotechrabbit GmbHNeuendorfstr. 24a16761 Hennigsdorf , [email protected]: +49 3302 20754 1 0Fax: +49 3302 20754 1 1

Commercial Register: Neuruppin (HRB 9381 NP)Managing Director: Dr. Bernd HaasePrincipal Office: HennigsdorfTax Office: OranienburgVAT no./USt-Id: DE 279969868Tax no.: 05310604935

Scientists around the world are working to leap ahead of diseases and to improve our lives with innovation. Aside from brilliant minds and relentless passion, the success of science depends on the quality of the materials used.

biotechrabbit is determined to offer the best products and services to those who lead the way in research. The valued relationships with our partners and customers drive us to exceed current limitations with flexibility, innovation and highly customized solutions to match their specific requirements. Each member of our team of highly engaged scientists, experienced managers and talented business developers aims to facilitate our partners and customers to leap and lead progress in the life science.

Our way of doing business combines the passion and pure curiosity of excellent researchers with the agile spirit of true entrepreneurs.

biotechrabbit

leap and lead

• Products made in Germany

• Diagnostic-grade quality

• OEM, custom and bulk

• PCR enzymes and mixes

• Antibody services

• Nucleic acid purification

• Cell-free protein synthesis

• mRNA for therapeutics

• Lyophilization services

About biotechrabbit

Contact Us

Ordering:Tel: +49 3302 207541 0Fax: +49 3302 207541 1 [email protected]

OEM and Antibody Service:Tel: +49 3302 207541 0Fax: +49 3302 207541 [email protected]

Technical Support:Tel: +49 3302 207541 2 Tel: +49 3302 207541 0Fax: +4903302 207541 1 [email protected]

Sales:Tel: +49 3302 207541 0Fax: +49 3302 207541 1 [email protected]

Collaboration and Partnership:Tel: +49 3302 207541 [email protected]

Career Opportunities:Tel :+49 3302 207541 [email protected]

3Bulk amounts and custom formulations available

www.biotechrabbit.com

Diagnostic Services and Custom Solutions 7 Enzymes for Diagnostics and Contract Manufacturing 7 Human Serum and Plasma for Diagnostics 8

Antibody Services 9 Monoclonal Antibody Development and Production 10 Polyclonal Antibody Development and Production 11

Standard PCR 13 Taq DNA Polymerase recombinant 14 Taq DNA Polymerase convenient 15 PCR Master Mix 15 Green Taq DNA Polymerase 16 Green PCR Master Mix 16 Red Taq DNA Polymerase 17 Red PCR Master Mix 17 MgCl2 18 PCR Enhancer 18 PCR Grade Water 18

Hot-Start PCR 19 Hot Start Taq DNA Polymerase 20 UPstart™ Taq Antibody 20 Hot Start Taq DNA Polymerase, convenient 21 Hot Start PCR Master Mix 21 Green Hot Start DNA Polymerase 22 Green Hot Start PCR Master Mix 22 Red Hot Start DNA Polymerase 23 Red Hot Start PCR Master Mix 23

Direct PCR, Blood and Crude Sample PCR 24 DirectUP™ PCR Kit 24

High-Fidelity and Long-Range PCR 25 Pfu DNA Polymerase 26 Pfu PCR Master Mix 26 Long And High Fidelity DNA Polymerase 27 Long Range PCR Master Mix 27

Reverse Transcription and RT-PCR 28 RevertUP™ Reverse Transcriptase 29 One Step RT-PCR Kit 29 MMuLV Reverse Transcriptase 30 RNase Inhibitor 30 cDNA Synthesis Kit 31

Quantitative Real-Time PCR 33 QPCR Green Master Mix 34 QPCR Probe Master Mix 34 QRT-PCR Green One Step Kits 35 QRT-PCR Probe One Step Kits 36

Nucleotides 37 dNTP and dNTP/dUTP Sets and Mixes 37

Product Overview

4

Bulk amounts and custom formulations available

Nucleic Acid Purification 38 DNA Cleanup GenUP™ PCR/Gel Cleanup Kit 40 GenUP™ Exo SAP PCR Cleanup Kit 40 GenUP™ PCR Cleanup Kit 41 GenUP™ Gel Extraction Kit 41 Plasmid Isolation GenUP™ Plasmid Kit 42 GenUP™ Plasmid Plus Kit 42 GenUP™ Plasmid Midi Kit 43 Total DNA Isolation GenUP™ Bacterial gDNA Kit 43 GenUP™ gDNA Kit 44 GenUP™ Mycobacteria gDNA Kit 44 GenUP™ Blood DNA Kit 45 GenUP™ Plant DNA Kit 45 Total RNA Isolation GenUP™ Total RNA Kit 46 GenUP™ Micro RNA Kit 46 GenUP™ Blood RNA Kit 47 GenUP™ Plant RNA Kit 47 UPzol™ RNA Isolation Solution 48 Virus DNA and RNA Isolation GenUP™ Virus RNA/DNA Kit 48 GenUP™ Virus RNA Kit 49 Deparaffinization and Bisulfite Conversion GenUP™ FFPE Paraffin Removal Solution 49 GenUP™ Bisulfite Complete Kit 50 GenUP™ Bisulfite Kit 50

Electrophoresis 51 DNA Electrophoresis Ladders and Loading Dye 51 Protein Electrophoresis Ladders 52

Enzymes for Molecular Biology 53 phi29 DNA Polymerase 54 StranDisplace™ Thermostable DNA Polymerase 54 T4 DNA Ligase, Rapid 55 Shrimp Alkaline Phosphatase 55 Heat Labile Uracil–DNA Glycosylase 56 Uracil–DNA Glycosylase 56 Heat Labile dsDNase 57 Proteinase K 57 T7 RNA Polymerase 58

Product Overview

5

Product OverviewProduct Overview

Cell-free Protein Synthesis 60

Cell-free Protein Synthesis Kits 61 E. coli Protein Synthesis Kits RTS 100 E. coli HY Kit 62 RTS 500 Proteomaster E. coli HY Kit 62 RTS 9000 E. coli HY Kit 63 EasyXpress E. coli Kit 63 RTS 100 E. coli Fab Kit 64 RTS 100/500 E. coli Disulfide Kits 64 Eukaryotic Protein Synthesis Kits RTS 100 Wheat Germ Kit 65 RTS 100/500 Wheat Germ CECF Kits 65 RTS 100 Insect Disulfide Kit 66 RTS 100 Insect Membrane Kit 66 RTS Supplements RTS Amino Acid Sampler 67 RTS Solubility Enhancer Set 67 RTS DnaK Supplement 68 RTS GroE Supplement 68

Protein Synthesis for Site-Directed Labeling 69 RTS 100 Site-Specific Label Kit 69

Linear Templates and Vectors for Cell-Free Protein Synthesis 70 Linear Templates for Cell-free Protein Synthesis RTS Linear Template Kit Plus 71 RTS Wheat Germ LinTempGen Set, His6-tag 71 EasyXpress Linear Template Kit Plus 72 RTS Linear Template Fab Kit 72 RTS 100 E. coli LinTempGen Set, His-tag 73 Vectors for Cell-free Protein Synthesis RTS pIX4.0 Insect Vector 73 RTS pIX3.0 Vector 74 RTS pIVEX Wheat Germ His6-tag Vector Set 74 RTS pIVEX E. coli His-tag, 2nd Gen. Vector Set 75 EasyXpress pIX3.0 Vector 75

Protein Purification His-Tag Purification Kit 76 His-Tag Purification Mag-Bead Kit 76

High Affinity His-tag Binding 77 Tris- NTA Amine / Biotin 77

www.biotechrabbit.com6

Diagnostic Services and Custom Solutions

biotechrabbit innovates, develops and manufactures products with superior performance for diagnostics, life science research and applied markets. We specialize in the production of enzymes for molecular biology and provide high-quality products with top-class availability.Most biotechrabbit products are available in bulk or as custom formulations. In close collaboration with our customers, we develop new enzymes, reagents and antibodies.

• Custom enzymes for diagnostics

• Contract manufacturing

• Antibody development

• Lyophilization Services

• OEM and Private Label

biotechrabbit‘s collaborative approach ensures reliability

Diagnostic-grade quality

Production according to diagnostic requirements• Audited quality according to ISO 9001 and 13485• Different manufacturing sites

Contract manufacturing

Large-scale enzyme and antibody production• Production in E. coli, eukaryotic and insect cells• Enzyme modeling, cloning and expression optimization

OEM and private label

Products tailor-made to your needs

• Custom formulation, individual packaging and labeling• Choice of high-quality molecular biology kits or enzymes

Antibody services

Mono- and polyclonal antibody development• Protein, peptide, or sequence as starting material• Antiserum, hybridomas or up to kg-scale mAbs

Cell-free synthesis platform

Protein and mRNA production• Fast and economic production for screening• Site-specific protein labeling or drug conjugates

Get in touch with biotechrabbit and start your project.

Phone: +49 3302 20754 1 0 Fax: +49 3302 20754 1 1 [email protected]

Lyophilization

Reagent stability at room temperature

• Freeze-dried PCR enzymes and mixes• Lyophilization service or glycerol-free enzymes

Bulk amounts and custom formulations available 7

Statistic Human Serum Panels and Matrix Selections

Statistic Human Serum Panel BR4000101

BR4000102

Panel of 100 Panel of 1000

100 × 500 µl Serum, normal donors1000 × 500 µl Serum, normal donors

Package contentCat. no. Size

FEATURES• High-quality true off-the-clot serum from normal donors in statistical distribution• Variation of serum in defined matrices• High ethical standards confirmed by ethic committees and informed donor consent• Full documentation for serum processing, donor origin, age and gender

• Diagnostic test development, optimization and validation• Biomarker identification

APPLICATIONS

DESCRIPTIONbiotechrabbit Human Serum is true off-the-clot serum in full statistical variation from donors representing a standard European population. The matrix selection contains single-donor serum from one donation in defined coagulants or anticoagulants. High ethical standards are confirmed by ethic committees and informed donor consent, according to international ethical and legal regulations. Documentation of age, gender and donor origin is provided, as well as specifications for blood collection and processing.High-quality serum panels or matrix selections ensure efficient and reliable development of diagnostic tests, including biomarker identification, assay optimization and validation.In addition to standard serum from “normal” donors, custom products can be provided for disease panels or defined panels according to donor origin in or outside Europe, age, gender, or disease state, serological tests, clinical documentation, special sample collection or other specific demands.

biotechrabbit provides highest quality human serum and plasma from normal donors for the following application areas:• Biomarker identification• Diagnostic test development and validation• Diagnostic kit production and QC

Compliance with ethical and legal regulations is guaranteed and complete documentation enables full traceability. The products are available as highly statistically significant panels as well as in bulk amounts and large pools.

Custom offers are designed to fulfill the specific testing needs of diagnostic test generation. Disease states with defined parameters are provided on request to serve all custom evaluation needs.

• Safe — full compliance with ethical and

legal regulations

• Reliable — highest quality serum and plasma

for consistent results

• Flexible — custom production

- Normal and disease state human biomaterial - Special parameters (high lipids, fasting, etc.) - Gender, age, origin of donor - Collection time, processing and analysis

Human Serum and Plasma for Diagnostics

Human Serum Matrix Selection BR4000201

BR4000202

4 Matrices

7 Matrices

500 µl each: Serum off the clot, K3EDTA, Sodium citrate, Lithium heparine serum500 µl each: Serum off the clot, K2EDTA, K3EDTA, Sodium citrate, Lithium heparin, Ammonium heparin, Serum by separator


Human Plasma Bulk and Pool

Human Bulk PlasmaBR4000301 300 ml 300 ml CPD Plasma from single

female or male donation


FEATURES• High-quality plasma pools from female or male donors• CPD plasma frozen only once to retain highest biomarker activity• Large batch quantities available• Stringent ethical standards and full documentation of processing and donor origin, age and gender

• Diagnostic test production and QC• Standard samples included in test kits

APPLICATIONS

DESCRIPTIONbiotechrabbit Human CPD Plasma is collected in single donations and directly frozen to ensure highest biomarker activity. This method allows a choice of male or female donors as well as custom mixes. Plasma Pools are available from female, male or mixed gender donors. Stringent ethical standards are confirmed by ethic committees and informed donor consent, according to international ethical and legal regulations. Documentation for age, gender and donor origin is provided as well as specifications on blood collection and processing.

High-quality plasma bulk and serum pools are available in large quantities for diagnostic test kit production and quality control as well as for standards to be included in the kits.In addition to plasma from “normal” donors, custom products can be provided for disease states. Large batches can be created according to defined specifications, such as donor origin, age, gender and disease state, serological tests, clinical documentation, special sample collection method other specific demands.

Human Plasma Pool FemaleBR4000401 BR4000402

100 ml

500 ml

100 ml CPD Plasma Pool, normal female donors 500 ml CPD Plasma Pool, normal female donors

Human Plasma Pool Mixed GenderBR4000601 BR4000602

100 ml

500 ml

100 ml CPD Plasma Pool, mixed normal gender (1:1)500 ml CPD Plasma Pool, mixed normal gender (1:1)

biotechrabbit‘s exceptional quality antibody development and high-capacity antibody production services are provided by qualified specialists who have been working in the field for more than 10 years. More than 1800 hybridomas have already been produced.

Large-scale production facilities, fermentation in serum- or protein-free medium and established purification schemes guarantee highest antibody purity. Endotoxin-free antibodies with cGMP-compliant documentation can be supplied for preclinical trials.

• Short project times — hybridomas or

antiserum in 5–6 weeks

• Flexible start— 500 µg to 1.5 mg protein,

peptide or protein sequence

• Best performance — highest purity of

monoclonal antibodies

• No delay — project start on short notice

Antibody Services

Human Plasma Pool MaleBR4000501

BR4000502

100 ml

500 ml

100 ml CPD Plasma Pool, normal male donors 500 ml CPD Plasma Pool, normal male donors


9

Monoclonal Antibody Development and Production

Features Description

• Monoclonal antibody development• Peptide-selection service

Peptide synthesis• Peptide-selection service included• Three mg peptide synthesised required for immunization and ELISA

BR3231001 Peptide synthesis for monoclonal antibody development

• Peptide or small protein• Monoclonal antibody development

Includes:• Conjugation of peptide or small protein to LPH or BSA protein carrier• Note: Peptide design to be discussed upfront with biotechrabbit

BR3232001 Conjugation to protein carrier

• Efficient immunization method• 70% of B lymphocytes are antigen specific

Immunization of three mice:• Protein antigen provided by customer: 500 µg protein (>1 mg/ml in phosphate buffer)• Alternative: immunization with peptide or project start with protein sequenceDuration: 17 days

BR3300101 Immunization of 3 Balb/c mice

• Fusion of B lymphocytes with myeloma cells• Selection of fused cells

Includes:• Preparation of lymph nodes (from three immunized mice)• Fusion with myeloma cell line P3-X63-Ag8• Distribution to 360 wells on culture plates• Cultivation and selection of fused cellsDuration: 10 days

BR3300201 Cell-fusion-producing hybridomas

• Screening and selection of clones• Expansion of positive clones

Includes:• ELISA screening of 360 oligoclones• Verification by ELISA, western blot analysis or immunoprecipitation• Selection and expansion of up to 24 positive clonesDuration: 2 weeks

BR3300301 Hybridoma screening and expansion

• Two subcloning rounds• Delivery of the final hybridoma cell lines

Includes:• First and second subcloning of one positive oligoclone• Verification by ELISA, western blot analysis or immunoprecipitation• Delivery of the final hybridoma cell linesDuration: 2 weeks for each subcloning

BR3300401 Hybridoma subcloning

• Highest purity standards• Production scale up to 1 kg

Cell line from hybridoma generation by biotechrabbit or customer to deliver hybridoma cell line.Includes:• Adaptation to serum-free media• Cell-line test for mycoplasma• Cultivation in serum-free or protein-free media• Purification by protein A/G affinity chromatography• Quality control and delivery of purified antibodyDuration:• Approximately 5 weeks for cell-line adaptation and mycoplasma test• Two weeks for cultivation to produce up to 40 mg• Two weeks for purification and quality control

BR3330601–6 Monoclonal antibody production, 5–1000 mg

• From oligoclone or subclone• Up to 50 µg antibody per clone

Includes:• Cultivation of oligoclone or subclone• Purification of 50 ml supernatant via affinity column• Antibody concentration and buffer exchange (PBS, Tris, or custom buffer)• Delivery of up to 50 µg antibody (depending on clone productivity)Duration: 2 weeks

BR3330501 Purified supernatant antibodies


Polyclonal Antibody Development and Production

Features Description

• High-quality antiserum• First serum delivery after 5 weeks

For two rabbits, protein antigen provided by customer (1.5 mg protein; >1 mg/ml)Includes:• Pre-immune sera, first immunization and boosts• Week 4: Bleeding, ELISA tests of sera and pre-sera (ELISA available only if protein is suitable for testing)• Week 5: Delivery of 1.5 ml pre-sera, approximately 20 ml antisera (each) and ELISA reports• Week 6: Customer confirms final bleeding or orders extension of immunization (BR3320104)• Week 7: Final bleeding• Week 8: Delivery of approximately 50 ml antiserum (each)

BR3320101 Polyclonal Rabbit Antiserum — Protein Antigen

• For polyclonal antibody development• Peptide-selection service

Peptide synthesis:• Peptide-selection service included• Price includes 15 mg of a standard 15 amino acid peptide, purity >80%• Final price depends on sequence, length, quantity and purity• Quantity calculated according to immunization protocol and additional services ordered• Customer receives remaining peptide at the end of service

BR3231002 Peptide synthesis for polyclonal antibody development

• High-quality antiserum• Peptide synthesis and conjugation to carrier

Peptide synthesis:• Peptide-selection service included• Synthesis of 15–20 mg peptide, purity >80%• Conjugation of peptide to protein carrier (LPH or BSA)• Customer receives remaining peptide at the end of serviceImmunization of 2 rabbits:• Pre-immune sera, first immunization and boosts• Week 4: Bleeding, ELISA tests of sera and pre-sera (ELISA only available if protein is suitable for testing)• Week 5: Delivery of 1.5 ml pre-sera, approximately 20 ml antisera (each) and ELISA reports• Week 6: Customer confirms final bleeding or orders extension of immunization (BR3320104)• Week 7: Final bleeding• Week 8: Delivery of approximately 50 ml antiserum (each)

BR3320102 Polyclonal Rabbit Antiserum — Peptide Antigen

• Guaranteed titer after immunization• Full refund, if titer is not attained

Must be ordered with protein or peptide immunization protocols, BR3320101/02 (protein from one batch of >90% purity)Including:• Protein pre-test in SDS gel• Antibody titer guarantee of at least 1:25,000 after protein immunization and 1:10,000 after peptide immunization (recognition of the antigen in an ELISA)• Full refund if guaranteed titer is not attained• Animal selection after pre-immune sera testing• Delivery of approximately 50 ml antisera (each)• Immunization schemes: over 5 weeks with protein antigen and over 6 weeks with peptide antigen, extension up to 3 months, if titer is not reached

BR3320103 Antibody Titer Guarantee

• Additional 20 ml high-quality antiserum• Delay of final bleeding

For one rabbit, using the same antigen as the initial immunization.Extension delays the final bleeding by 4 weeks and can be repeated up to 2 years.Includes:• Immunization boost• 4 weeks animal keeping• Approximately 20 ml serum

BR3320104 Extension of Immunization (4 weeks)



NOTES



Standard PCR

Standard PCR products include a highly active Taq DNA Polymerase that is suitable for demanding applications, such as diagnostics and routine amplification. PCR master mixes, which contain all necessary PCR reagents, simplify PCR setup. Colored, high-density PCR buffers allow PCR products to be loaded directly onto gels after cycling. Colored storage buffers allow easy identification of mixes containing Taq DNA polymerase during reaction setup. Taq DNA polymerase allows targets of 3–5 kb to be routinely amplified under standard or fast cycling conditions. In addition, an adenine is added to the 3' end of the product, allowing TA cloning.

• Diagnostic-grade quality — top performance

for demanding application

• Convenient — master mixes and colored

enzymes make your job easier

• Efficient — abundant DNA ready for

downstream applications

Product Main featureMain PCR application

Simplified setup Flexibility

Direct gel loading

Colored enzyme

Taq DNA Polymerase recombinant, 5 U/µl

Taq DNA Polymeraseconvenient, 5 U/µl

PCR Master Mix, 2×

Green Taq DNA Polymerase, 5 U/µl

Green PCR Master Mix, 2×

Red Taq DNA Polymerase, 5 U/µl

Red PCR Master Mix, 2×

50 mM MgCl2

5× PCR Enhancer

PCR Grade Water

MgCl2 supplied in a separate vial for maximum flexibility

Reaction buffer contains standard MgCl2 concentration

Premixed PCR reagents; just add template, primers and water

Green-colored buffer for direct loading onto gel after PCR

Premixed PCR including green-colored buffer for direct loading onto gel after the PCR

Red-colored storage buffer for identification of mixes containing the enzyme

Premixed PCR reagents including red-colored buffer for identification of mixes containing the enzyme

Flexible optimization of Mg2+ concentration

Improved PCR specificity and efficiency

Ultrapure, sterile filtrated, contamination-free

Standard and demanding


Routine, high-throughput

Standard with direct gel loading

Routine, high-throughput with direct gel loading

Standard with enzyme identification

Routine, high-throughput with enzyme identification




Bulk amounts and custom formulations available 13 Bulk amounts and custom formulations available

14

Taq DNA Polymerase recombinant, 5 U/µl

• Routine and applied PCR up to 5 kb• RT-PCR• TA cloning

APPLICATIONS

FEATURES• High product yields and robustness in a wide application range• Highest quality utilized in molecular diagnostics and research• Exceptionally pure Taq DNA Polymerase for routine and demanding PCR applications

DESCRIPTIONbiotechrabbit Taq DNA polymerase is a first-choice enzyme for all routine and molecular diagnostics PCR applications . The exceptional quality and purity of the enzyme ensures the highest performance that is required by the diagnostic industry and research labs. The polymerase is suitable for standard and fast PCR applications ensuring high product yields from various templates with targets of up to 5 kb in size.

biotechrabbit Taq DNA polymerase is a thermostable, highly processive 5'→3' DNA polymerase that has low 5'→3' exonuclease activity and lacks 3'→5' exonuclease (proofreading) activity. The latter allows incorporation of modified nucleotides.

The enzyme also exhibits deoxynucleotidyl transferase activity that results in the addition of extra A overhang at the 3'ends of PCR products, allowing easy cloning of PCR products into vectors with T overhangs.

BR0100101

BR0100102

BR0100103

100 U

500 U

2500 U

20 µl Taq DNA Polymerase1.8 ml 10×Reaction Buffer1.5 ml 50 mM MgCl2

100 µl Taq DNA Polymerase2 × 1.8 ml 10× Reaction Buffer1.5 ml 50 mM MgCl2

5 × 100 µl Taq DNA Polymerase10 × 1.8 ml 10× Reaction Buffer5 × 1.5 ml 50 mM MgCl2


QUALITY CONTROL — ensuring highest performance

Functional assayHuman genomic DNA was amplified using Taq DNA Poly-merase and specific primers to produce a distinct band of 750 bp.

Self-priming activityStandard PCR is carried out without primers, using Taq DNA Polymerase and human genomic DNA. No products were amplified.

Exonuclease assayLinearized lambda/HindIII fragments are incubated with Taq DNA Polymerase in a 50 µl reaction mixture for 4 h at 37°C. No degradation of DNA was observed.

Endonuclease assayLambda DNA is incubated with of Taq DNA Polymerase in a 50 µl reaction mixture for 4 h at 37°C. No degradation of DNA was observed.

Nick assay Supercoiled plasmid DNA is incubated with Taq DNA Polymerase in a 50 µl reaction mixture for 4 h at 37°C. No conversion of covalently closed circular DNA to nicked DNA was detected.

E. coli genomic DNA contamination assayA sample of denatured Taq DNA Polymerase is analyzed with specific primers for the 16S rRNA gene in qPCR for the presence of contaminating E. coli genomic DNA. The detection limit is < 1 copy E. coli genomic DNA per unit Taq DNA Polymerase. No E. coli genomic DNA was detectable.

Unit DefinitionOne unit is defined as the amount of Taq DNA Polymerase required to catalyze the incorporation of 10 nmol dNTP into acid-insoluble form in 30 min at 72°C in the presence of the reaction buffer.


15

Taq DNA Polymerase convenient, 5 U/µl

• Routine PCR up to 5 kb• RT-PCR• TA cloning

APPLICATIONS

FEATURES• Optimal concentration of MgCl2 included in Reaction Buffer• High product yields and robustness in a wide application range• Exceptionally pure Taq DNA Polymerase for both routine and demanding PCR applications

DESCRIPTIONbiotechrabbit Taq DNA Polymerase is a first-choice ther-mostable DNA polymerase for all routine PCR applica-tions, ensuring high product yields from various templates for targets up to 5 kb in size.

Our Taq DNA Polymerase convenient version provides optimal MgCl2 concentration in the Reaction Buffer, saving time during reaction setup.

biotechrabbit Taq DNA Polymerase is a thermostable, highly processive 5'→3' DNA polymerase that has low 5'→3' exonuclease activity and lacks 3'→5' exonuclease (proofreading) activity. The latter activity allows incorpora-tion of modified nucleotides into DNA.

The enzyme also exhibits deoxynucleotidyltransferase activity, resulting in the addition of an extra A overhang at the 3' ends of PCR products, allowing easy cloning of PCR products into vectors with T overhangs.

PCR Master Mix, 2×

FEATURES• Optimized PCR Master Mix for minimal hands-on and fast setup• Exceptionally pure Taq DNA Polymerase and highest quality dNTPs• High product yields and robustness in a wide application range

APPLICATIONS• Routine and high-throughput PCR up to 5 kb• TA cloning

DESCRIPTIONbiotechrabbit PCR Master Mix is a perfect choice for fast reaction setup that reduces the time required for calcu-lation and pipetting and eliminates the need for buffer optimization. It is designed for routine, high-throughput PCR amplification of 0.2–5 kb DNA targets. The 2× Master Mix contains highly purified recombinant biotechrabbit Taq DNA Polymerase, extremely high-quality dNTPs and optimized PCR buffer; thus, only template DNA, PCR primers and PCR-grade water need to be added.

biotechrabbit Taq DNA Polymerase is a thermostable, highly processive 5'→3' DNA polymerase that has low 5'→3' exonuclease activity and lacks 3'→5' exonuclease (proofreading) activity. The latter activity allows incorpora-tion of modified nucleotides into DNA.

The enzyme also exhibits deoxynucleotidyltransferase activity that results in the addition of an extra A overhang at the 3' ends of PCR products, allowing easy cloning of PCR products into vectors with T overhangs.

BR0100701

BR0100702

BR0100703

100 U

500 U

2500 U

20 µl Taq DNA Polymerase1.8 ml 10× Reaction Buffer with MgCl2

100 µl Taq DNA Polymerase2 × 1.8 ml 10× Reaction Buffer with MgCl2

5 × 100 µl Taq DNA Polymerase10 × 1.8 ml 10× Reaction Buffer with MgCl2


BR0100201

BR0100202

BR0100203

100 rxn of 50 µl 500 rxn of 50 µl 2500 rxn of 50 µl

2 × 1.25 ml PCR Master Mix 10 × 1.25 ml PCR Master Mix 50 × 1.25 ml PCR Master Mix



16

Green Taq DNA Polymerase, 5 U/µl

• PCR and immediate gel analysis• Routine PCR up to 5 kb• TA cloning

APPLICATIONS

FEATURES• Green Reaction Buffer formulation for direct loading on the gel• High product yields and robustness in a wide application range• Exceptionally pure Taq DNA Polymerase for both routine and demanding PCR applications

DESCRIPTIONbiotechrabbit Green Taq DNA Polymerase is a first-choice thermostable DNA polymerase for routine PCR applica-tions that allows direct electrophoresis without the need to add loading buffer and ensures high product yields from various templates.

The 5× Reaction Buffer of Green Taq DNA Polymerase is recommended for any amplification reaction that will be visualized by agarose gel electrophoresis. The buffer contains two dyes (blue and yellow) that separate during electrophoresis, allowing the migration progress to be monitored. Reactions assembled with 5× Reaction Buffer have sufficient density for direct loading onto agarose gels.

biotechrabbit Taq DNA Polymerase is a thermostable, highly processive 5'→3' DNA polymerase that has low 5'→3' exonuclease activity and lacks 3'→5' exo-nuclease (proofreading) activity. The enzyme also exhibits deoxyribonucleotidyl transferase activity that results in the addition of an extra A overhang at the 3' ends of PCR products.

Green PCR Master Mix, 2×

FEATURES• Optimized Green PCR Master Mix for fast setup and direct loading on the gel• Exceptionally pure Taq DNA Polymerase and highest quality dNTPs• High product yields and robustness in a wide application range

APPLICATIONS• High-throughput PCR and immediate gel analysis• Routine PCR up to 5 kb• TA cloning

DESCRIPTIONbiotechrabbit Green PCR Master Mix is a perfect choice for a fast reaction setup that reduces the time required for calculation and pipetting and eliminates the need for buffer optimization. Additionally the special formulation allows reactions onto be loaded onto an agarose gel directly after amplification without a separate step for adding loading dye.

Green PCR Master Mix contains two dyes (blue and yellow) that separate during electrophoresis, allowing the migration progress to be monitored. Reactions with the Green PCR Master Mix have sufficient density for direct loading onto agarose gels. Green Reaction Buffer also allows mixtures containing the enzyme to be identified.

The Green PCR Master Mix contains highly purified recombinant biotechrabbit Taq DNA Polymerase, extremely high-quality dNTPs and optimized PCR buffer; thus, only template DNA, PCR primers and PCR-grade water need to be added.

BR0100301

BR0100302

BR0100303

100 U

500 U

2500 U

20 µl Taq DNA Polymerase2 × 1.8 ml 5× Green Reaction Buffer1.5 ml 50 mM MgCl2

100 µl Taq DNA Polymerase4 × 1.8 ml 5× Green Reaction Buffer1.5 ml 50 mM MgCl2 5 × 100 µl Taq DNA Polymerase20 × 1.8 ml 5× Green Reaction Buffer5 × 1.5 ml 50 mM MgCl 2


BR0100401

BR0100402

BR0100403

100 rxn of 50 µl 500 rxn of 50 µl 2500 rxnof 50 µl

2 × 1.25 ml Green PCR Master Mix 10 × 1.25 ml Green PCRMaster Mix 50 × 1.25 ml Green PCR Master Mix



17

Red Taq DNA Polymerase, 5 U/µl

• Simplified PCR setup with easy identification of the reaction mixtures that contain the enzyme• Routine PCR up to 5 kb• TA cloning

APPLICATIONS

FEATURES• Red Taq DNA Polymerase Storage Buffer for identifying mixtures containing the enzyme• High product yields and robustness in a wide application range• Exceptionally pure Taq DNA Polymerase for both routine and demanding PCR applications

DESCRIPTIONbiotechrabbit Red Taq DNA Polymerase is a first-choice enzyme ensuring high product yields and simplified PCR setup. This red-colored enzyme formulation performs in all respects like colorless polymerases. Additionally, reaction mixtures containing the enzyme are easily identified due to the red-colored enzyme Storage Buffer.

Red Taq DNA Polymerase is a thermostable, highly processive 5'→3' DNA polymerase that has low 5'→3' exonuclease activity and lacks 3'→5' exonuclease (proofreading) activity. The enzyme also exhibits deoxynucleotidyltransferase activity that results in the addition of an extra A overhang at the 3' ends of PCR, products allowing easy cloning of PCR products into vectors with T overhangs.

Red PCR Master Mix, 2×

FEATURES• Optimized Red PCR Master Mix for fast and easy setup• Exceptionally pure Taq DNA Polymerase and highest quality dNTPs• High product yields and robustness in a wide application range

APPLICATIONS• High-throughput PCR setup with easy visualization of the reaction mixtures• Routine PCR up to 5 kb• TA cloning

DESCRIPTIONbiotechrabbit Red PCR Master Mix is a perfect choice for simplified reaction setup that reduces the time required for calculation and pipetting and eliminates the need for buffer optimization. Additionally the colored formulation provides the possibility to identify the reactions which already contain the enzyme and dNTPs.

This red-colored master mix performs in all respects like colorless PCR master mixes and ensures high product yields from various templates. It contains highly purified recombinant biotechrabbit Taq DNA Polymerase, extremely high-quality dNTPs and optimized PCR buffer; thus, only template DNA, PCR primers and PCR-grade water need to be added.

BR0100801

BR0100802

BR0100803

100 U

500 U

2500 U

20 µl Red Taq DNA Polymerase 1.8 ml 10× Reaction Buffer1.5 ml 50 mM MgCl2

100 µl Red Taq DNA Polymerase 2 × 1.8 ml 10× Reaction Buffer1.5 ml 50 mM MgCl2

5 × 100 µl Red Taq DNA Polymerase 10 × 1.8 ml 10× Reaction Buffer5 × 1.5 ml 50 mM MgCl2


BR0100601

BR0100602

BR0100603

100 rxn of 50 µl 500 rxn of 50 µl 2500 rxnof 50 µl

2 × 1.25 ml Red PCR Master Mix 10 × 1.25 ml Red PCR Master Mix 50 × 1.25 ml Red PCR Master Mix



18

MgCl2

• Optimization of PCR buffer conditionsAPPLICATIONS

FEATURES• Flexibility in PCR optimization• Improved PCR performance

DESCRIPTIONbiotechrabbit MgCl2 is ideally suited for optimization of PCR conditions. For convenience and flexibility, it is provided as 50 mM solution.

PCR Enhancer

FEATURES• Increased specificity and efficiency• Improved results with GC-rich templates

APPLICATIONS• Optimize PCR performance in demanding applications• Sensitive, low-background amplification

DESCRIPTIONbiotechrabbit PCR Enhancer is a unique PCR additive that improves sensitivity, resulting in increased PCRefficiency and reduced background. PCR Enhancer is ideal for difficult templates (e.g., GC-rich) and complex reactions, such as low-abundance templates and multiplex PCR.

BR1900101 6 ml 4 × 1.5 ml 50 mM MgCl2


BR1900201 6 ml 4 × 1.5 ml 5× PCR EnhancerPackage contentCat. no. Size

PCR Grade Water

FEATURES• Ultrapure and sterile filtrated• DNase-, RNase- and protease-free, no DNA contamination

APPLICATIONS• Ready-to-use water for standard and enhanced PCR and qPCR• Buffer preparation and enzyme dilutions

DESCRIPTIONbiotechrabbit PCR Grade Water is ultrapure and sterile filtered water for use in all PCR applications. It is tested for bacterial and mammalian DNA contamination and for DNases, RNases and proteases as well. Aliquots of 1.5 ml support an easy and contamination-free assay setup.

BR1900301 15 ml 10 × 1.5 ml PCR Grade WaterPackage contentCat. no. Size


19

Hot-Start PCR

Hot-start enzymes are completely inactive during room-temperature setup and become active only after heating, reducing random primer annealing and unspecific amplification that can occur with standard PCR enzymes. The fidelity of hot-start PCR is the same as for standard PCR, and targets of 3–5 kb are routinely amplified. Like standard Taq DNA polymerase, hot-start enzymes add an adenine to the 3' end of the product, allowing TA cloning.

Hot-start PCR master mixes, which contain all necessary PCR reagents, simplify PCR setup. Colored, high-density hot-start PCR buffers allow PCR products to be loaded directly onto gels after cycling. Colored storage buffers allow easy identification of mixes containing Hot Start Taq DNA Polymerase during reaction setup.

• Optimized — excellent enzyme performance

for enhanced applications

• Specific and sensitive — hot start for

better results

• Convenient — component choices that

make your job easier

• Fast — master mixes for less pipetting

Product Main featureMain PCR application

Simplified setup Flexibility

Direct gel loading

Colored enzyme

Hot Start Taq DNA Polymerase, 5 U/µl

UPstart™ Taq Antibody, 1 mg/ml

Hot Start Taq DNA Polymerase convenient, 5 U/µl

Hot Start PCR Master Mix, 2×

Green Hot Start DNA Polymerase, 5 U/µl

Green Hot Start PCR Master Mix, 2×

Red Hot Start DNA Polymerase, 5 U/µl

Red Hot Start PCR Master Mix, 2×

Antibody-based hot start polymerase for improved PCR specificity and yield

Hot-start using Taq DNA polymerases

Antibody-based hot start supplied with PCR buffer containing Mg2+

Antibody-based hot start with premixed PCR reagents; just add template, primers and water

Antibody-based hot start with green-colored buffer for direct loading onto gels after PCR

Antibody-based hot start with premixed PCR reagents and green-colored buffer for direct loading onto gels after PCR

Antibody-based hot start with red-colored buffer for easy identification of mixes containing enzyme

Antibody-based hot start with premixed PCR reagents and red-colored buffer

High-sensitivity, high-specificity



High-throughput, high-specificity, high-sensitivity

High-specificity, high-sensitivity, direct gel loading

High-throughput, high-specificity, high-sensitivity, direct gel loading

High-sensitivity, high-specificity, identification of enzyme mix

High-throughput, high- specificity, iden tification of enzyme mix


20

Hot Start Taq DNA Polymerase, 5 U/µl

• Hot-start PCR up to 5 kb• Amplification of low-copy-number targets• RT-PCR and TA cloning

APPLICATIONS

FEATURES• High PCR specificity and sensitivity• Exceptionally pure Hot Start Taq DNA Polymerase for sensitive PCR applications and high yields• Antibody-based Hot Start for fast polymerase activation

DESCRIPTIONbiotechrabbit Hot Start Taq DNA Polymerase is a first-choice hot-start PCR enzyme for all demanding PCR applications. The enzyme ensures high product yields with low background and without primer–dimer formation and nonspecific priming.

The Hot Start Taq DNA Polymerase is inactive during reaction setup due to the bound antibody which is quickly released at elevated temperatures, ensuring the enzyme is active only during PCR. There is no need for prolonged heating or denaturation steps.

The optional use of 5× PCR Enhancer improves PCR results in many cases, including impure template or low template abundance.

UPstart™ Taq Antibody, 1 mg/ml

FEATURES• Inhibition of >95% Taq activity at 45°C• 200 ng UPstart Taq Antibody are blocking 1 U Taq DNA polymerase• Exceptionally pure— made in cell culture

APPLICATIONS• Thermolabile inhibition of Taq DNA polymerases• Convenient hot-start PCR setup at room temperature• Fast polymerase activation with the first PCR denaturation step

DESCRIPTIONbiotechrabbit UPstart Taq Antibody is an ultra-puremonoclonal antibody against the Taq DNA polymerase. It is produced in cell culture to ensure highest quality.

The antibody can be used with highly efficient Taq DNA polymerases, provides an excellent method for “hot start” PCR and enhances PCR specificity and sensitivity. PCR hot start prevents the formation of primer–dimers and nonspecific amplification and allows convenient PCR setup at room temperature.

In the first denaturation step of the thermal cycling, the UPstart Antibody becomes nonfunctional and the active Taq DNA polymerase is released. This antibody-mediated hot-start method is significantly more convenient to use than other hot-start methods. Polymerase reactivation using this antibody is faster than with methods using chemically inhibited polymerases.

BR0200101

BR0200102

BR0200103

100 U

500 U

2500 U

20 µl Hot Start Taq DNA Polymerase1.8 ml 10× Reaction Buffer1.5 ml 50 mM MgCl2

500 µl 5× PCR Enhancer 100 µl Hot Start Taq DNA Polymerase2 × 1.8 ml 10× Reaction Buffer1.5 ml 50 mM MgCl2

1.5 ml 5× PCR Enhancer 5 × 100 µl Hot Start Taq DNA Polymerase10 × 1.8 ml 10× Reaction Buffer5 × 1.5 ml 50 mM MgCl2

5 × 1.5 ml 5× PCR Enhancer


BR1200101

BR1200102

BR1200103

100 µg (500 U) 250 µg (1250 U) 1000 µg (5000 U)

100 µl UPstart Taq Antibody 250 µl UPstart Taq Antibody

1000 µl UPstart Taq Antibody



21

Hot Start Taq DNA Polymerase convenient, 5 U/µl

• High-specificity hot-start PCR up to 5 kb• Amplification of low-copy-number targets• RT-PCR and TA cloning

APPLICATIONS

FEATURES• Optimal concentration of MgCl2 included in Reaction Buffer• High-sensitivity and robustness in a wide application range• Exceptionally pure Hot Start Taq DNA Polymerase for both routine and demanding PCR applications

DESCRIPTIONbiotechrabbit Hot Start Taq DNA Polymerase is a first-choice hot-start PCR enzyme for all demanding PCR applications. The enzyme ensures high product yields with low background and without primer–dimer formation and nonspecific priming.Our Hot Start Taq DNA Polymerase convenient version provides optimal MgCl2 concentration in the Reaction Buffer, saving time during reaction setup.The Hot Start Taq DNA Polymerase is inactive during reaction setup due to the bound antibody which is quickly released at elevated temperatures, ensuring the enzyme is active only during PCR. There is no need for prolonged heating or denaturation steps. The optional use of 5× PCR Enhancer improves PCR results in many cases, including impure template or low template abundance.

Hot Start PCR Master Mix, 2×

FEATURES• Highest PCR sensitivity without prolonged reactivation• Optimized PCR Master Mix for minimal hands-on and fast setup• Exceptionally pure Hot Start Taq DNA Polymerase and highest quality dNTPs

APPLICATIONS• High-specificity and high-throughput hot-start PCR up to 5 kb• Amplification of low-copy-number targets• TA cloning

DESCRIPTIONbiotechrabbit Hot Start PCR Master Mix is a perfect choice for a fast reaction setup that reduces the time required for calculation and pipetting and eliminates the need for buffer optimization. It is designed for low-background, high-throughput PCR of 0.2–5 kb DNA targets.

The 2x Hot Start PCR Master Mix contains pure biotechrabbit Hot Start Taq DNA Polymerase, extremely high-quality dNTPs and optimized PCR buffer; thus, only template, PCR primers and PCR-grade water are added.

The Hot Start Taq DNA Polymerase is inactive during reaction setup due to the bound antibody, which is quickly released at elevated temperatures, ensuring the enzyme is active only during PCR. There is no need for prolonged heating or denaturation steps. The hot start minimizes primer–dimers and miss-priming.


BR0200601

BR0200602

BR0200603

100 U

500 U

2500 U

20 µl Hot Start Taq DNA Polymerase1.8 ml 10× Reaction Buffer with MgCl2

500 µl 5× PCR Enhancer 100 µl Hot Start Taq DNA Polymerase2 × 1.8 ml 10× Reaction Buffer with MgCl2

1.5 ml 5× PCR Enhancer5 × 100 µl Hot Start Taq DNA Polymerase10 × 1.8 ml 10× Reaction Bufferwith MgCl2



BR0200201

BR0200202

BR0200203

100 rxn of 50 µl

500 rxn of 50 µl

2500 rxnof 50 µl

2 × 1.25 ml Hot Start PCR Master Mix 500 µl 5× PCR Enhancer 10 × 1.25 ml Hot Start PCR Master Mix 1.5 ml 5× PCR Enhancer50 × 1.25 ml Hot Start PCR Master Mix 5 × 1.5 ml 5× PCR Enhancer



22

Green Hot Start DNA Polymerase, 5 U/µl

• High-sensitivity PCR and immediate gel analysis• High-specificity hot-start PCR up to 5 kb• Amplification of low-copy-number targets• RT-PCR and TA cloning

APPLICATIONS

FEATURES• Green Reaction Buffer formulation for direct loading on the gel right after PCR• High PCR specificity and sensitivity• Hot Start Taq DNA Polymerase for demanding sensitive PCR applications and high yields

DESCRIPTIONbiotechrabbit Green Hot Start DNA Polymerase is a first-choice hot-start PCR enzyme for all demanding PCR applications. The enzyme ensures high product yields with low background and without primer –dimer formation and nonspecific priming that allows direct electrophoresis and ensures high product yields from various templates.The 5× Reaction Buffer of Green Hot Start Taq DNA Poly-merase is recommended for any amplification reaction that will be visualized by agarose gel electrophoresis. The Buffer contains two dyes (blue and yellow) that separate during electrophoresis, allowing the migration progress to be mo-nitored. Reactions assembled with the 5× Reaction Buffer have sufficient density for direct loading onto agarose gels. The optional use of 5x PCR Enhancer improves PCR results in many cases, including impure template or low template abundance.

Green Hot Start PCR Master Mix, 2×

• High-throughput hot-start PCR and immediate gel analysis• High-specificity hot-start PCR up to 5 kb• Amplification of low-copy-number targets• TA cloning

APPLICATIONS

FEATURES• Optimized Green Hot Start PCR Master Mix for fast setup and direct loading on the gel• Exceptionally pure Hot Start Taq DNA Polymerase and highest quality dNTPs• High product yields and robustness in a wide application range

DESCRIPTIONbiotechrabbit Green Hot Start PCR Master Mix is a perfect choice for a fast reaction setup that reduces the time required for calculation and pipetting and eliminates the need for buffer optimization. It is designed for low-background, high-throughput PCR amplification of 0.2–5 kb DNA targets. Additionally the special formulation allows reactions to be loaded directly onto gels after amplification without adding additional loading dye.

Green Hot Start PCR Master Mix contains two dyes (blue and yellow) that separate during electrophoresis, allowing migration progress to be monitored. Reactions with Green Hot Start PCR Master Mix have sufficient density for direct loading onto agarose gels.


BR0200501

BR0200502

BR0200503

100 rxn of 50 µl

500 rxn of 50 µl

2500 rxnof 50 µl

2 × 1.25 ml Green Hot Start PCR Master Mix500 µl 5× PCR Enhancer10 × 1.25 ml Green Hot Start PCR Master Mix 1.5 ml 5× PCR Enhancer50 × 1.25 ml Green Hot Start PCR Master Mix 5 × 1.5 ml 5× PCR Enhancer


BR0200701

BR0200702

BR0200703

100 U

500 U

2500 U

20 µl Hot Start Taq DNA Polymerase1.8 ml 5× Green Reaction Buffer1.5 ml 50 mM MgCl2

500 µl 5× PCR Enhancer100 µl Hot Start Taq DNA Polymerase2 × 1.8 ml 5× Green Reaction Buffer1.5 ml 50 mM MgCl2

1.5 ml 5× PCR Enhancer5 × 100 µl Hot Start Taq DNA Polymerase10 × 1.8 ml 5× Green Reaction Buffer5 × 1.5 ml 50 mM MgCl2




23

Red Hot Start DNA Polymerase, 5 U/µl

• High-throughput simplified PCR setup with easy identification of the reaction mixtures that contain the enzyme• High-specificity hot-start PCR up to 5 kb• Amplification of low-copy-number targets• TA cloning

APPLICATIONS

FEATURES• Red Hot Start DNA Polymerase storage buffer for identifying mixtures containing the enzyme• Exceptionally pure Hot Start Taq DNA Polymerase for both routine and demanding PCR applications• Highest PCR specificity and sensitivity without prolonged reactivation

DESCRIPTIONbiotechrabbit Red Hot Start DNA Polymerase is a first-choice hot-start enzyme ensuring high product yields with low background and simplified PCR setup. The red-colo-red enzyme performs in all respects like colorless hot-start polymerase, ensures low-background and high-specificity amplification. Additionally, re action mixtures containing the enzyme are easily identified due to the red-colored enzyme Storage Buffer.The Hot Start Taq DNA Polymerase is inactive during reaction setup due to the bound antibody, which is quickly released at elevated temperatures, ensuring the enzyme is active only during PCR, reducing primer–dimer formation and miss-priming. The optional use of 5× PCR Enhancer improves PCR results in many cases, including impure template or low template abundance.

BR0200301

BR0200302

BR0200303

100 U

500 U

2500 U

20 µl Red Hot Start DNA Polymerase1.8 ml 10× Reaction Buffer1.5 ml 50 mM MgCl2

500 µl 5× PCR Enhancer100 µl Red Hot Start DNA Polymerase2 × 1.8 ml 10× Reaction Buffer1.5 ml 50 mM MgCl2

1.5 ml 5× PCR Enhancer5 × 100 µl Red Hot Start DNA Polyme-rase10 × 1.8 ml 10× Reaction Buffer5 × 1.5 ml 50 mM MgCl2



Red Hot Start PCR Master Mix, 2×

FEATURES• Optimized Red Hot Start PCR Master Mix for highest sensitivity and easy setup• Exceptionally pure Hot Start Taq DNA Polymerase and highest quality dNTPs• High product yields and robustness in a wide application range

APPLICATIONS• High-throughput PCR setup with easy visualization of the reaction mixtures• Hot-start PCR up to 5 kb• Amplification of low-copy-number targets• TA cloning

DESCRIPTIONbiotechrabbit Red Hot Start PCR Master Mix is a perfect choice for simplified hot-start PCR setup that reduces the time required for calculation and pipetting and eliminates the need for buffer optimization. Additionally, the colored formulation allows the reactions that contain the enzyme and dNTPs to be identified.


The 2× Red Hot Start PCR Master Mix contains pure biotechrabbit Hot Start Taq DNA Polymerase, extremely high-quality dNTPs and optimal PCR buffer; thus, only template, PCR primers and PCR-grade water are added.

BR0200401

BR0200402

BR0200403

100 rxn of 50 µl

500 rxn of 50 µl

2500 rxnof 50 µl

2 × 1.25 ml Red Hot Start PCR Master Mix500 µl 5× PCR Enhancer10 × 1.25 ml Red Hot Start PCR Master Mix5 × 1.5 ml 5× PCR Enhancer50 × 1.25 ml Red Hot Start PCR Master Mix5 × 1.5 ml 5× PCR Enhancer



24

Direct PCR, Blood and Crude Sample PCR

The DirectUP DNA Polymerase is an inhibitor-resistant thermophilic polymerase that enables amplification from crude samples without prior DNA template purification. The time- and cost-saving technique allows fast detection of targets directly from blood, soil, food samples and tissue homogenates.

• Fast — for direct amplification of unpurified

targets

• Robust — resistant to inhibitors in blood and

crude samples

DirectUP™ PCR Kit

• Direct PCR from blood, soil or other crude samples without template purification

APPLICATIONS

FEATURES• Inhibition-resistant hot-start DNA Polymerase for direct PCR from blood or crude samples• Excellent performance in a wide application range• Tolerates up to 40% whole blood, inhibitors in soil, foods and intercalating dyes

DESCRIPTIONbiotechrabbit DirectUP PCR Kit contains an engineered hot-start DNA polymerase providing excellent results without the need for template purification. The enzyme retains robust PCR function in the presence of 40% whole blood and is resistant to inhibitors present in complex samples, such as serum, soil, inhibitory foods and fluorescent dyes.

A series of mutations provides an effective hot-start function, ensuring minimized background and eliminating primer–dimer formation. The enzyme lacks both 5'→3' and 3'→5' exonuclease activities.

The enzyme exhibits deoxynucleotidyl transferase activity that results in the addition of an extra A overhang at the 3' ends of PCR products allowing easy cloning of PCR products into vectors with T overhangs.

BR0100901

BR0100902

BR0100903

100 rxn of 25 µl 400 rxn of 25 µl

1000 rxn of 25 µl

100 µl DirectUP DNA Polymerase1 ml 5× DirectUP Reaction Buffer400 µl DirectUP DNA Polymerase2 × 1 ml 5× DirectUP Reaction Buffer1 ml DirectUP DNA Polymerase5 × 1 ml 5× DirectUP Reaction Buffer



25

High-Fidelity and Long-Range PCR

Pfu DNA Polymerase provides PCR accuracy that is approximately 10 times higher than Taq DNA Polymerase, and is suitable for demanding cloning, sequencing and expression applications.Long-range PCR kits provide an optimized combination of Taq DNA polymerase and a proofreading DNA polymerase for longer PCR products with higher accuracy than is possible with the Taq enzyme alone. Long-range enzymes amplify templates of up to 30–40 kb with high fidelity. PCR Enhancer is supplied for better results with GC-rich ampli-cons. High-fidelity and long-range products are not recommended for use with dUTP.

• Optimized — excellent enzyme performance

for enhanced applications

• Accurate — high-fidelity polymerase

• Reliable — polymerase for large amplicons

• Fast — master mixes for less pipetting

during setup

Product Main featureMain PCR application Fidelity*

Amplicon length

Simplified setup

Colored enzyme

Pfu DNA Polymerase, 2.5 U/µl

Pfu PCR Master Mix, 2×

Long and High Fidelity DNA Polymerase, 2.5 U/µl

Long Range PCR Master Mix, 2×

Ten times more accurate than Taq DNA Polymerase

Includes Pfu DNA Polymerase and high-fidelity PCR reagents

DNA polymerases for long-range and high-fidelity PCR

DNA polymerases and reagents for long-range and high-fidelity PCR

High-fidelity,demanding PCR

High-throughput, high-fidelity

Long-range, GC-rich templates

High-throughput, long-range

~10×higher

~10×higher

~4× higher

~4× higher

3 kb

3 kb

>30 kb

>30 kb


* vs. Taq DNA Polymerase

26

Pfu DNA Polymerase, 2.5 U/µl

• High-fidelity PCR• Generation of PCR products for blunt cloning• Site directed mutagenesis

APPLICATIONS

FEATURES• Accurate PCR for demanding applications• Approximately ten times higher accuracy than that of Taq DNA Polymerase• Proof-reading for increased fidelity

DESCRIPTIONbiotechrabbit Pfu DNA Polymerase is a highly purified ther-mostable recombinant proofreading DNA polymerase. Pfu DNA Polymerase exhibits approximately 10 times higher accuracy than Taq DNA polymerase and amplifies targets up to 3–4 kb in size.

The enzyme catalyzes template-dependent nucleo-tide polymerization in the 5'→3' direction. Additionally the 3'→5' exonuclease (proofreading) activity corrects nucleotide incorporation errors, thereby increasing fidelity and accuracy of DNA polymerization. The enzyme has no 5'→3' exonuclease activity and no detectable reverse transcriptase activity and produces blunt-end PCR products.

For the most demanding applications, the supplied 5× PCR Enhancer can be optionally used for improving results when using templates with GC-rich sequences and complex structures.

Pfu PCR Master Mix, 2×

• High-throughput, high-fidelity PCR• Generation of PCR products for blunt cloning

APPLICATIONS

FEATURES• Optimized Pfu PCR Master Mix for minimal hands-on and fast setup• Pure Pfu DNA Polymerase and highest quality dNTPs• Approximately ten times higher accuracy than that of Taq DNA Polymerase for accurate PCR in demanding applications

DESCRIPTIONbiotechrabbit Pfu PCR Master Mix is a perfect choice for fast, high-fidelity PCR setup that reduces the time required for calculation and pipetting and eliminates the need for buffer optimization. It is designed for routine high-throughput, high-fidelity amplification of targets up to 3–4 kb in size.

The 2× Pfu PCR Master Mix contains Pfu DNA Polymerase, extremely high-quality dNTPs and optimized PCR buffer; thus, only template, PCR primers and PCR-grade water are added.

For the most demanding applications, the supplied 5× PCR Enhancer can be optionally be used to improve results when using templates with GC-rich sequences and complex structures.

Pfu DNA Polymerase exhibits approximately 10 times higher accuracy compared to Taq DNA polymerase. Pfu DNA Polymerase produces blunt-end PCR products suitable for blunt cloning.

BR0300201

BR0300202


2 × 1.25 ml Pfu PCR Master Mix 500 µl 5× PCR Enhancer10 × 1.25 ml Pfu PCR Master Mix 1.5 ml 5× PCR Enhancer


BR0300101

BR0300102

100 U

500 U

40 µl Pfu DNA Polymerase 500 µl 10× Pfu Reaction Buffer500 µl 5× PCR Enhancer200 µl Pfu DNA Polymerase 2 × 1.2 ml 10× Pfu Reaction Buffer1.5 ml 5× PCR Enhancer



27

Long and High Fidelity DNA Polymerase, 2.5 U/µl

• Long-range PCR up to 40 kb• Amplification of GC-rich templates

APPLICATIONS

FEATURES• High-productivity, long-range PCR • Increased fidelity for accurate amplification of GC-rich templates• Polymerase mix for high yield and short cycle times

DESCRIPTIONbiotechrabbit Long and High Fidelity DNA Polymerase is a first-choice for amplification of targets up to 40 kb in size with higher accuracy than that of Taq DNA polymerase.

This specially designed blend of thermophilic polymerases is well suited for amplification of targets that are GC-rich and have complex structures.

For the most demanding applications, the supplied 5× PCR Enhancer can be optionally used to improve results when using templates with GC-rich sequences and complex structures.

Long and High Fidelity DNA Polymerase produces a mix-ture of A-tailed and blunt-end PCR products. It is advisable to blunt products before cloning into blunt-end vectors.

BR0300301

BR0300302

100 U

500 U

40 µl Long and High Fidelity PCR Enzyme Mix500 µl 10× HF Reaction Buffer500 µl 5× PCR Enhancer200 µl Long and High Fidelity PCR Enzyme Mix2 × 1.2 ml 10× HF Reaction Buffer1.5 ml 5× PCR Enhancer


Long Range PCR Master Mix, 2×

FEATURES• Optimized Long Range PCR Master Mix for minimal hands-on and fast setup• Mix of pure polymerases and highest quality dNTPs for high yield and short cycle times• Increased fidelity for accurate amplification of GC-rich templates

APPLICATIONS• High-throughput, long-range PCR up to 40 kb• Amplification of GC-rich templates

DESCRIPTIONbiotechrabbit Long Range PCR Master Mix is a perfect choice for fast reaction setup for long-range PCR that reduces the time required for calculation and pipetting and eliminates the need for buffer optimization. It is designed for amplification of targets up to 40 kb in size. The master mix works well with GC-rich templates and amplifies DNA with a higher fidelity than that of Taq DNA polymerase.

The Long Range PCR Master Mix contains a blend of ther-mophilic polymerases, extremely high-quality dNTPs and optimized PCR buffer; thus, only template, PCR primers and PCR-grade water are added.

For the most demanding applications, the supplied 5× PCR Enhancer can be optionally used to improve results when using templates with GC-rich sequences and complex structures.

Long Range PCR Master Mix produces a mixture of A-tailed and blunt-end PCR products. It is advisable to blunt products before cloning into the blunt-end vector.

BR0300401

BR0300402

100 rxn of 50 µl

500 rxn of 50 µl

2 × 1.25 ml Long Range PCR Master Mix 500 µl 5×PCR Enhancer10 × 1.25 ml Long Range PCR Master Mix 1.5 ml 5× PCR Enhancer



28

Reverse Transcription and RT-PCR

RevertUP Reverse Transcriptase, which is a proprietary modification of the MMuLV reverse transcriptase, provides most efficient cDNA synthesis without RNase H activity, allowing successful synthesis of cDNAs of greater than 14 kb in length.One-step RT-PCR kits provide optimized reagents, including Hot Start Taq DNA Polymerase, for performing both reverse-transcription and amplification reactions in one tube.The Moloney-Murine Leukemia Virus (MMuLV) reverse transcriptase is a classic RNA-dependent DNA polymerase. The exceptional purity of the biotechrabbit MMuLV enzyme ensures excellent results in demanding applications. A special RNase Inhibitor prevents RNA templates from degradation by pervasive RNases.

• High performance — exceptional purity for

demanding applications

• Convenient — excellent efficiency at high

temperatures

• Reliable — RNase Inhibitor for protecting

your RNA

Product Main PCR application Main feature

RevertUP™ Reverse Transcriptase

One Step RT-PCR Kit

MMuLV Reverse Transcriptase

RNase Inhibitor

cDNA Synthesis Kit

Reliable synthesis of ≥14 kb cDNA, two-step RT-PCR

One-step RT-PCR

cDNA synthesis, two-step RT-PCR

Prevention of RNA degradation by RNases

First-strand cDNA synthesisfor RT-PCR and qPCR

A proprietary MMuLV reverse transcriptase engineered by point mutations in polymerase and RNase H domains ensures efficient cDNA synthesis with no RNase H activity, allowing successful synthesis of ≥14 kb cDNA.

A blend of efficient thermostable reverse transcriptase and proprietary Ribonuclease Inhibitor ensures high cDNA yields.Unique Hot Start Taq DNA Polymerase in a mix with high-quality dNTPs and PCR enhancers allows sensitive, low-background amplification.

Exceptionally pure reverse transcriptase supplied with reaction buffer for standard applications.

A recombinant, non-competitive inhibitor of pancreatic-type ribonucleases, including RNase A, RNase B and RNase C.

Excellent yields at temperatures up to 55°C and superior performance in demanding applications, including templates with a high degree of secondary structures


29

RevertUP™ Reverse Transcriptase, 200 U/µl

• First-strand cDNA synthesis• Generation of labeled cDNA• RNA analysis by primer extension• cDNA library construction• RT-PCR

APPLICATIONS

FEATURES• Improved performance for synthesis of long cDNAs (≥ 14 kb)• Excellent efficiency at high temperatures up to 55 °C• High sensitivity for cDNA synthesis from few copies of template

DESCRIPTIONbiotechrabbit RevertUP Reverse Transcriptase is a proprietary MMuLV reverse transcriptase engineered by point mutations in polymerase and RNase H domains. This ensures efficient cDNA synthesis and eliminates RNase H activity, allowing successful synthesis of ≥14 kb cDNA.

Reverse transcriptase is a DNA polymerase which uses RNA as a substrate and exhibits no measurable proofreading 3'→5' exonuclease function. This enzyme performs cDNA synthesis by extending a DNA primer annealed to an RNA template; it can also make copies of single-stranded DNA templates.

BR0400301

BR0400302

10000 U

50000 U

50 µl RevertUP Reverse Transcriptase1.25 ml 5× RevertUP Buffer250 µl RevertUP Reverse Transcriptase4 × 1.25 ml 5× RevertUP Buffer


One Step RT-PCR Kit

FEATURES• Efficient thermostable Reverse Transcriptase and proprietary Ribonuclease Inhibitor providing high cDNA yields• Unique Hot Start Taq DNA Polymerase in a mix with high-quality dNTPs• PCR enhancers allowing sensitive low background amplification

APPLICATIONS• One-step RT-PCR• Virus detection• Amplification of GC-rich and complex templates

DESCRIPTIONbiotechrabbit One Step RT-PCR Kit provides an easy and efficient way to perform both reverse transcription of RNA and PCR amplification of cDNA in one step. Only RNA template, primers and PCR-grade water are added.

The 20× RT-RI Blend, which contains a blend of an efficient thermostable reverse transcriptase and a proprietary Ribonuclease Inhibitor, ensures high yields of cDNA.

The 2× One Step Mix, which contains unique Hot Start Taq DNA Polymerase, dNTPs, MgCl2 and stabilizers in an optimized buffer, provides high PCR product yields with minimal background even when using low-abundance templates and difficult targets. PCR enhancers included in the mix allow efficient amplification of complex templates including GC- or AT-rich sequences.

BR0400101

BR0400102

BR0400103


1.25 ml One Step Mix 125 µl RT-RI Blend2 × 1.25 ml One Step Mix 2 × 125 µl RT-RI Blend10 × 1.25 ml One Step Mix 10 × 125 µl RT-RI Blend



30

MMuLV Reverse Transcriptase, 200 U/µl

• First-strand cDNA synthesis• Generation of labeled cDNA• RNA analysis by primer extension

APPLICATIONS

FEATURES• Pure reverse transcriptase for cDNA synthesis• High yields of first-strand cDNA• High value for a fair price

DESCRIPTIONbiotechrabbit MMuLV Reverse Transcriptase is an exceptionally pure DNA polymerase which uses RNA as a substrate and exhibits no measurable proofreading 3'→5' exonuclease function. This enzyme performs cDNA synthesis by extending a DNA primer annealed to an RNA template; it can also make copies from a single-stranded DNA templates.

The enzyme is purified from a recombinant E. coli strain carrying the MMuLV reverse transcriptase gene.

BR0400201

BR0400202

10000 U

50000 U

50 µl MMuLV Reverse Transcriptase500 µl 10× MMuLV RT Buffer250 µl MMuLV Reverse Transcriptase1.5 ml 10× MMuLV RT Buffer


RNase Inhibitor, 40 U/µl

FEATURES• Exceptionally pure proprietary Ribonuclease Inhibitor for demanding RNA applications• Active under variety of reaction conditions used for work with RNA• Prevention of RNA from degradation by RNase A, RNase B and RNase C

APPLICATIONS• In vitro transcription, translation• cDNA synthesis• RNA purification and storage

DESCRIPTIONbiotechrabbit RNase Inhibitor is an acidic, 52 kDa protein that is a potent non-competitive inhibitor of pancreatic-type ribonucleases such as RNase A, RNase B and RNase C. The enzyme is purified from a recombinant E. coli strain carrying a fusion protein from the porcine RNase Inhibitor gene with a proprietary 22.5 kDa protein tag.

BR0400901BR0400902

2500 U 10000 U

62.5 µl RNase Inhibitor 250 µl RNase Inhibitor




cDNA Synthesis Kit

• First-strand cDNA synthesis for RT-PCR and qPCR• Gene expression profiling• RNA labeling and primer extension• cDNA library construction

APPLICATIONS

FEATURES• Highly efficient synthesis of long cDNAs (≥ 14 kb)• Excellent yields at temperatures up to 55°C• High sensitivity reverse transcription from low abundance template• Superior performance in demanding applications, including templates with a high degree of secondary structure

DESCRIPTIONbiotechrabbit cDNA Synthesis Kit provides superior components that ensure efficient first-strand cDNA synthesis from mRNA or total RNA templates. biotechrabbit RevertUP™ Reverse Transcriptase, a proprietary MMuLV reverse transcriptase, carries point mutations in the polymerase and RNase H domains, eliminating RNase H activity and ensuring highly efficient reverse transcription with increased thermostability.

biotechrabbit RNase Inhibitor is a potent non-competitive inhibitor of RNases. The combination of highly efficient cDNA synthesis, effective RNase inhibition and out-standingly pure nucleotide allows high yields of cDNAs of more than 14 kb. For greater application flexibility, random hexamer primers, allowing all RNAs in the reaction to be used as templates, and an oligo (dT) primer, allowing synthesis of cDNA from only poly(A) tailed mRNA, are included.

BR0400401

BR0400402

BR0400403

10 rxn of 20 µl

25 rxn of 20 µl

125 rxn of 20 µl

10 µl RevertUP Reverse Transcriptase 100 µl 5× cDNA Synthesis Buffer20 µl M dNTP Mix5 µl RNase Inhibitor10 µl Hexamer, 5 µl Oligo (dT) Primers1.5 ml PCR Grade Water25 µlRevertUP Reverse Transcriptase100 µl 5× cDNA Synthesis Buffer50 µl dNTP Mix12.5 µl RNase Inhibitor25 µl Hexamer, 12.5 µl Oligo (dT) Primers1.5 ml PCR Grade Water125 µl RevertUP Reverse Transcriptase500 µl 5× cDNA Synthesis Buffer250 µl dNTP Mix62.5 µl RNase Inhibitor125 µl Hexamer, 62.5 µl Oligo (dT) Primers5 × 1.5 ml PCR Grade Water



NOTES

33

Quantitative Real-Time PCR

biotechrabbit QPCR Green, Probe and One-Step QRT-PCR Master Mixes are first-choice products for fast and easy real-time PCR setup. They contain all reagents required for real-time PCR and are designed to achieve excellent results in reaction effi ciency, correlation coefficient and slope. Intercalating-dye-based mixes, such as QPCR Green Master Mix, allow nonspecific detection of double-stranded DNA and are faster and more cost-effective than probe-based mixes.Probe-based mixes, such as QPCR Probe Master Mix, can be used specifically or universally. However, more effort is required to design and synthesized specific probes.biotechrabbit QPCR and QRT-PCR products can be used with the fast and standard modes as well as for multiplex PCR.The mixes are available with passive reference dyes Rox™ (carboxy-X-rhodamine) or fluorescein to enable the correction of errors caused by the instrument.

• High performance — rapid extension rate

for early Ct values

• Sensitive — increased limit of detection

for quicker results

• Compatible — use on any real-time

PCR platform

• Flexible — for standard and fast cycling

conditions

Reference dye Company Instrument

Low concentration

ROX

High concentration

ROX

Fluorescein

Analytica JenaApplied BiosystemsBio-Rad

CepheidEppendorfIlluminaQIAGENRoche Applied ScienceAgilentTakaraTechne

Analytica JenaApplied Biosystems

CepheidEppendorfIlluminaQIAGENRoche Applied ScienceTakaraTechne

Bio-Rad

qTower7500, 7500 FAST, Viia™ 7iCycler®, MyiQ™, iQ™ 5, Opticon™, Opticon 2, Chromo4™, MiniOpticon™ , CFX96™, CFX384™Smartcycler®Mastercycler® ep REALPLEX®, Mastercycler REALPLEX 2SEco™Rotor-Gene® 3000, 6000, QLightcycler® 480, Lightcycler NanoMx4000P®, Mx3000P®, Mx3005P®Cycler Dice™Quantica®

qTower7000, 7300, 7700, 7900, 7900HT, 7900HT FAST, StepOne™, StepOnePlus™SmartcyclerMastercycler ep REALPLEX, Mastercycler REALPLEX 2SEcoRotor-Gene 3000, 6000, QLightcycler 480, Lightcycler NanoCycler DiceQuantica

iCycler, MyiQ, iQ 5

Instrument compatibility for passive reference dyes


34

QPCR Green Master Mix, 2×

QPCR Green Master Mix LRox, 2×BR0500301 BR0500302 BR0500304


1 ml QPCR Green Master Mix LRox5 × 1 ml QPCR Green Master Mix LRox25 × 1 ml QPCR Green Master Mix LRox


QPCR Green Master Mix HRox, 2×BR0500401

BR0500402 BR0500404


1 ml QPCR Green Master Mix HRox5 × 1 ml QPCR Green Master Mix HRox25 × 1 ml QPCR Green Master Mix HRox

QPCR Green Master Mix Fluorescein, 2×BR0501201

BR0501202

BR0501203


1 ml QPCR Green Master Mix Fluorescein5 × 1 ml QPCR Green Master Mix Fluorescein25 × 1 ml QPCR Green Master Mix Fluorescein

• qPCR based on intercalating dye• Standard and fast cycling• Compatible on all real-time PCR platforms

APPLICATIONS

FEATURES• Non-PCR-inhibiting intercalating dye• Highest sensitivity for increased limit of detection• Rapid extension rate for early Ct values

DESCRIPTIONbiotechrabbit QPCR Green Master Mixes can be used to quantify any DNA template including genomic, cDNA and viral sequen ces. Extremely low-copy-number targets can be detected specifically with high efficiency in standard and fast cycling. The mixes use a proprietary intercala ting dye which does not inhibit PCR. Hot-start function prevents formation of primer–dimers and nonspecific products leading to improved reaction sensitivity and specificity. For greater flexibility, master mixes with fluorescein and ROX at high (HRox) or low (LRox) concentration are available.

QPCR Probe Master Mix, 2×

QPCR Probe Master Mix, 2×BR0501301

BR0501302

BR0501304


1 ml QPCR Probe Master Mix

5 × 1 ml QPCR Probe Master Mix

25 × 1 ml QPCR Probe Master Mix


QPCR Probe Master Mix LRox, 2×BR0500501

BR0500502 BR0500504


QPCR Probe Master Mix HRox, 2×BR0500601

BR0500602

BR0500604


1 ml QPCR Probe Master Mix LRox5 × 1 ml QPCR Probe Master Mix LRox25 × 1 ml QPCR Probe Master Mix LRox

1 ml QPCR Probe Master Mix HRox5 × 1 ml QPCR Probe Master Mix HRox25 × 1 ml QPCR Probe Master Mix HRox

• qPCR based on specific probes• Standard and fast cycling• For use on a wide range of probe technologies including Taqman®, Molecular Beacons® and Scorpion® probes

APPLICATIONS

FEATURES• Highest sensitivity for increased limit of detection• High efficiency in multiplex reactions• Rapid extension rate for early Ct values

DESCRIPTIONbiotechrabbit QPCR Probe Master Mixes can be used to quantify any DNA template including genomic, cDNA and viral sequen ces. Extremely low-copy-number targets can be detected specifically with high efficiency in standard and fast cycling. The mixes are compatible with many probe technologies. Hot-start function prevents formation of primer–dimers and nonspecific products leading to improved reaction sensitivity and specificity. For greater flexibility, master mixes without reference dye, with fluorescein and ROX at high (HRox) or low (LRox) concentration are available.


35

QRT-PCR Green One Step Kit, 2×

QRT-PCR Green OneStep Kit LRoxBR0500801 BR0500802

BR0500803


1 ml QPCR Green Mix LRox200 µl RTase with RNase Inhibitor3 × 1 ml QPCR Green Mix LRox3 × 200 µl RTase with RNase Inhibitor12 × 1 ml QPCR Green Mix LRox12 × 200 µl RTase with RNase Inhbitor


QRT-PCR Green OneStep Kit HRoxBR0500701

BR0500702

BR0500703

100 rxn of 20 µl


1 ml QPCR Green Mix HRox200 µl RTase with RNase Inhibitor3 × 1 ml QPCR Green Mix HRox3 × 200 µl RTase with RNase Inhibitor12 × 1 ml QPCR Green Mix HRox12 × 200 µl RTase with RNase Inhi-bitor

• One step QRT-PCR based on intercalating dye• For use with standard and fast q PCR platforms

APPLICATIONS

FEATURES• Highest sensitivity for increased limit of detection• Thermostable, extremely active reverse transcriptase (55°C) and advanced RNase inhibitor• Non-PCR inhibiting intercalating dye• Rapid extension rate for early Ct values

DESCRIPTIONbiotechrabbit QRT-PCR Green One Step Kits allow efficient cDNA synthesis and qPCR in a single tube. They can be used to quantify mRNA, total RNA and viral sequences. Extremely low-copy-number targets can be detected with high efficiency. Hot-start function prevents formation of primer–dimers and nonspecific products leading to improved reaction sensitivity and specificity.

QRT-PCR Green One Step Kits perform excellently in standard cycling conditions as well as in fast and ultra-fast cycling conditions. For greater flexibility, one-step kits with ROX at high (HRox) or low (LRox) concentration are available.

Uniform and reproducible results with QPCR Master MixEfficiency of the amplification of the E. coli 16S RNA gene using tenfold serial dilutions in duplicates from 4.2 ng to 4.2 pg in a QPCR Probe assay.


36

QRT-PCR Probe One Step Kit, 2×

QRT-PCR Probe OneStep KitBR0501101

BR0501102

BR0501103


1 ml QPCR Probe Mix200 µl RTase with RNase Inhibitor3 × 1 ml QPCR Probe Mix3 × 200 µl RTase with RNase Inhibitor12 × 1 ml QPCR Probe Mix12 × 200 µl RTase with RNase Inhibitor


QRT-PCR Probe OneStep Kit LRoxBR0500901

BR0500902

BR0500903


1 ml QPCR Probe Mix LRox200 µl RTase with RNase Inhibitor3 × 1 ml QPCR Probe Mix LRox3 × 200 µl RTase with RNase Inhibitor12 × 1 ml QPCR Probe Mix LRox12 × 200 µl RTase with RNase Inhibitor

QRT-PCR Probe OneStep Kit HRoxBR0501001

BR0501002

BR0501003


1 ml QPCR Probe Mix HRox200 µl RTase with RNase Inhibitor3 × 1 ml QPCR Probe Mix HRox3 × 200 µl RTase with RNase Inhibitor12 × 1 ml QPCR Probe Mix HRox12 × 200 µl RTase with RNase Inhibitor

FEATURES• Highest sensitivity for increased limit of detection• Thermostable, extremely active reverse transcriptase (55°C) and advanced RNase inhibitor• Rapid extension rate for early Ct values

• One-step RT-QPCR based on specific probes• For use with standard and fast qPCR platforms

APPLICATIONS

DESCRIPTIONbiotechrabbit QRT-PCR Probe One Step Kits are first-choice products for a fast and easy real-time PCR setup using RNA templates. They contain all reagents required for one-step QPCR and are designed to achieve excellent results in reaction efficiency, correlation coefficient and slope.To enable the use of the kit on qPCR platforms with different reference dye concentration requirements, three kit versions are available: a one-step kit containing no ROX and LRox and HRox versions containing ROX in high and low concentrations, respectively.QRT-PCR Probe One Step Kits use proprietary reverse transcriptase technology and buffer chemistry for efficient cDNA synthesis and QPCR in a single tube. They can be used to quantify any RNA template including mRNA, total RNA and viral sequences. Extremely low-copy-number targets can be detected with high efficiency. Hot-start function prevents primer–dimers and nonspecific products leading to improved reaction sensitivity and specificity. Kits also provide leading performance in fast and ultra-fast cycling conditions.


37

Nucleotides

dNTP Sets and Mixes

dNTP Set (4× 100 mM solutions)BR0600601

BR0600602

BR0600603

BR0600605

4 × 250 µl

4 × 1 ml

5 pack of 4 × 250 µl5 pack of 4 × 1 ml

250 µl 100 mM solutions each: dATP/dCTP/dGTP/dTTP1 ml 100 mM solutions each: dATP/dCTP/dGTP/dTTP5 × 250 µl 100 mM solutions each: dATP/dCTP/dGTP/dTTP 5 × 1 ml 100 mM solutions each: dATP/dCTP/dGTP/dTTP

Package contentCat. no. Size FEATURES• Exceptional quality dNTPs of >99% purity confirmed by HPLC• Free from DNA and PCR inhibitors• Consistent PCR results due to outstanding dNTPs stability

• Standard or hot-start PCR• Long-range and high-fidelity PCR• cDNA synthesis and RT-PCR• qPCR• Sequencing• DNA labeling

APPLICATIONS

DESCRIPTIONbiotechrabbit deoxynucleotide triphosphates are first-choice nucleotides for all PCR applications, including the most demanding, such as amplification of long targets (up to 40 kb), GC-rich templates, qPCR, cDNA synthesis, high-fidelity PCR, DNA labeling and sequencing.Advanced production technology ensures that deoxy-ribonucleotide triphosphates have >99% purity and outstanding stability, ensuring excellent performance and consistent, reliable results.For the maximum flexibility, nucleotides are available in sets and mixes of common concentrations.

dNTP/dUTP Set (4× 100 mM solutions)BR0600701 4 × 250 µl 250 µl 100 mM solutions each:

dATP/dCTP/dGTP/dTUP25 mM dNTP MixBR0600501BR0600502BR0600503

250 µl1 ml 5 ml

250 µl dNTP Mix (25 mM each)1 ml dNTP Mix (25 mM each)5 × 1 ml dNTP Mix (25 mM each)

10 mM dNTP MixBR0600201BR0600202BR0600204

250 µl1 ml 5 ml

250 µl dNTP Mix (10 mM each)1 ml dNTP Mix (10 mM each)5 × 1 ml dNTP Mix (10 mM each)

2 mM dNTP MixBR0600301BR0600302

1 ml5 ml

1 ml dNTP Mix (2 mM each)5 × 1 ml dNTP Mix (2 mM each)

dNTP/dUTP MixBR0600801

BR0600802

1 ml

5 ml

1 ml dNTP/dUTP Mix (2 mM dATP/dCTP/dGTP, 4 mM dUTP)5 × 1 ml dNTP/dUTP Mix (2 mM dATP/dCTP/dGTP, 4 mM dUTP)

biotechrabbit deoxynucleotide triphosphates are available as mixes containing all dNTPs or sets of individual dNTPs. Sets and mixes containing dUTP instead of dTTP are available for applications such as PCR carryover prevention. The outstanding purity of the nucleotides ensures excellent performance in the most demanding applications.

• Reproducible — exceptional purity for

excellent results

• Reliable — outstanding stability for

consistent PCR results


100 mM ATPBR0601001BR0601002

250 µl1 ml

250 µl ATP (100 mM4 × 250 µl ATP (100 mM))

100 mM dATPBR0600901BR0600902

250 µl1 ml

250 µl dATP (100 mM4 × 250 µl dATP (100 mM))


Nucleic Acid Purification

biotechrabbit nucleic acid purification kits are designed to be fast and convenient to use. Purified nucleic acids are suitable for standard and demanding applications. Universal kits provide reagents and protocols for purification from a wide range of starting materials.Mini filter based procedures allow easy handling, and proprietary buffer compositions ensure most efficient lysis, nucleic acid binding, washing and elution.

• Convenient — simple mini filter based

procedures to make your work easier

• Efficient — proprietary buffer compositions for

high yields of pure nucleic acid

Product Applications Starting material

DNA cleanup

GenUP PCR/Gel Cleanup Kit

GenUP Exo SAP PCR Cleanup Kit

GenUP PCR Cleanup Kit

GenUP Gel Extraction Kit

Plasmid isolation

GenUP Plasmid Kit

GenUP Plasmid Plus Kit

GenUP Plasmid Midi Kit

Total DNA isolation

GenUP Bacteria gDNA Kit

GenUP gDNA Kit

GenUP Mycobacteria gDNA Kit

GenUP Blood DNA Kit

GenUP Plant DNA Kit

PCR product cleanup and DNA extraction from agarose gels

PCR cleanup for sequencing with 100% product recovery

Mini Filter based PCR product cleanup

DNA extraction from agarose gels

Universal kit for isolating high- and low-copy-number plasmids from bacterial culture

Universal kit for isolating high- and low-copy-number plasmids from different volumes of bacterial culture

High- or low-copy plasmid isolation DNA from up to 50 ml bacterial cultures

Universal kit for genomic DNA isolation from bacterial cultures

Universal kit for genomic DNA isolation from various eukaryotic starting materials

Isolation of genomic DNA from mycobacteria

Optimized kit for DNA isolation from blood

Optimized kit for DNA isolation from plants

Amplification reactions and TAE or TBE agarose gels

Amplification reactions

Amplification reactions

TAE or TBE agarose gels

Gram-positive and Gram-negative bacteria




Tissues, rodent tails, paraffin-embedded samples, swabs, eukaryotic cells suspensions

Sputum sample, bronchoalveolar lavage or tissue biopsies

Fresh or frozen whole blood stabilized with EDTA or citrate

Fresh, frozen or dry plant materials


Product Applications Starting material

Total RNA isolation

GenUP Total RNA Kit

GenUP Micro RNA Kit

GenUP Blood RNA Kit

GenUP Plant RNA Kit

UPzol RNA Isolation Solution

GenUP Virus DNA/RNA Kit

GenUP Virus RNA Kit

GenUP FFPE Paraffin Removal Solution

GenUP Bisulfite Complete Kit

GenUP Bisulfite Kit

Universal kit for total RNA isolation from various sources

Isolation of snRNA, miRNA, siRNA, tRNA, rRNA and mRNA

Optimized kit for total RNA isolation from blood

Optimized kit for total RNA isolation from plants

Modified guanidine isothiocyanate/phenol method for total RNA isolation from different sources using scalable volumes

Optimized kit for simultaneous viral DNA and RNA isolation from eukaryotic samples

Optimized kit for viral RNA isolation from various eukaryotic samples

Removal of paraffin from formalin-fixed, paraffin-embedded (FFPE) sample

Isolation of genomic DNA, bisulfite conversion of unmethylated cytosines to uracils, subsequent desulfonation and DNA cleanup.

Bisulfite conversion of unmethylated cytosines to uracils for epigenomics studies

Tissue, eukaryotic cell suspensions, bacteria

Eukaryotic cells, tissue or biopsies, Gram-posi-tive and Gram-negative bacteria

Fresh or frozen whole blood stabilized with EDTA or citrate

Fresh, frozen or dry plant materials

Tissues and cell suspensions

Serum, plasma and other cell-free body fluids, supernatants from cell cultures, tissues, biopsies, cell cultures, swabs, paraffin-embedded samples

Serum, plasma, cell-free body fluids, cell culture supernatant, tissues, biopsies, cell cultures, swabs, paraffin-embedded samples

FFPE slices

Purified DNA, cells, tissue, FFPE samples, bronchial aspirates, swabs or body fluids

Purified DNA (1 pg to 10 µg)

Nucleic Acid Purification

• Flexible — purification for a range of sample

types with one kit

• Reliable — best results in downstream

applications

• Optimized — no toxic β-mercaptoethanol and

no DNase for RNA preparations

• Fast — PCR cleanup in only 3 min

Virus DNA and RNA isolation

Deparaffinization and bisulfite conversion

40

GenUP™ PCR/Gel Cleanup Kit

BR0700501BR0700502 BR0700503

10 preps 50 preps 250 preps

Solutions, mini filters and vialsSolutions, mini filters and vialsSolutions, mini filters and vials

Package content

• Fast purification of DNA from agarose gels• Fast purification of PCR products from amplification reactions

APPLICATIONS

FEATURES• Dual performance kit for both PCR product cleanup and DNA purification from agarose gels• Fast and simple procedure• High DNA recovery yields

DESCRIPTIONbiotechrabbit GenUP PCR/Gel Extraction Kit has been specially developed for a quick and easy cleanup or concentration of PCR fragments from reaction mixtures as well as extraction of DNA from both TAE and TBE agarose gels. The DNA is bound to a Mini Filter using a novel buffer, washed and then eluted in a separate tube. The purified DNA is ready to be used in all demanding molecular bio-logy applications, including restriction digestion, ligation, sequencing, transfection into mammalian cells and in vitro transcription.

Starting materialExtraction timeBinding capacityTypical yieldAverage purity

TAE or TBE agarose gel (up to 300 mg)Approximately 20 min> 20 µg DNA100 bp – 30 kb60–90%

GenUP™ Exo SAP PCR Cleanup Kit

FEATURES• Fast 15 min PCR cleanup without the need for spin columns• 100% recovery — no loss of PCR product• Simple process — just add Exonuclease 1 and Shrimp Alkaline Phosphatase to PCR mix• Easy scalability and automation

APPLICATIONS• PCR cleanup prior to sequencing or genotyping• Removal of nucleotides and primers from PCR reaction

DESCRIPTIONbiotechrabbit GenUP Exo SAP PCR Cleanup Kit is designed for quickly and easily cleaning up PCR amplifi-cation reactions, enabling their direct use in sequencing or genotyping. The kit provides a combination of Shrimp Alkaline Phosphatase (SAP) and Exonuclease 1 (Exo) for removing nucleotides and primers. Double-stranded DNA is not affected, ensuring 100% recovery of PCR products.

Both enzymes are active in PCR buffers and completely inactivated by heating for 10 min at 80°C: no further processing is required. Shrimp Alkaline Phosphatase dephosphorylates nucleotides and primers, and the 5' → 3' exonuclease activity of Exonuclease 1 degrades single-stranded primers.

Cat. no. Size


PCR mixtures (up to 50 µl)Approximately 3 min> 20 µg DNA60 bp – 30 kb60–95%

BR0701801

BR0701802

BR0701803

100 rnx

500 rnx

2000 rnx

100 µl Shrimp Alkaline Phosphatase100 µl Exonuclease 1500 µl Shrimp Alkaline Phosphatase500 µl Exonuclease 14 × 500 µl Shrimp Alkaline Phospha-tase4 × 500 µl Exonuclease 1


Gel cleanup

PCR cleanup


41

GenUP™ PCR Cleanup Kit

• Fast purification of PCR amplification reactionsAPPLICATIONS

FEATURES• Fast and convenient PCR cleanup procedure in only 3 min• Just bind and elute, no need for washing steps• High-purity DNA recovery for all demanding applications• High DNA recovery yields

DESCRIPTIONbiotechrabbit GenUP PCR Cleanup Kit has been developed for quick and easy cleanup or concentration of PCR fragments from reaction mixtures. High-yield PCR product with excellent quality is purified from the PCR mixtures in a simple two-step procedure. DNA is bound to a Mini Filter using a novel buffer and eluted. The need for a washing step has been eliminated, reducing hands-on-time. The procedure takes approximately 3 min, compared to 8 min required to other kits.

The purified DNA is ready for use in all demanding mole-cular biology applications, including restriction digestion, ligation and sequencing.

Starting material

Extraction timeBinding capacityTypical yieldAverage purity

Amplification reaction mixtures (up to 50 µl)Approximately 3 min> 20 µg DNA60 bp – 30 kb60–95%

GenUP™ Gel Extraction Kit

FEATURES• Fast and simple DNA extraction from agarose gels• High recovery yields• Excellent purity DNA for demanding applications

APPLICATIONS• Fast purification of DNA from TAE or TBE agarose gels

DESCRIPTIONbiotechrabbit GenUP Gel Extraction Kit has been specially developed for a quick and easy extraction of DNA from both TAE and TBE agarose gels. After few initial procedures, the DNA is bound to a Mini Filter using a novel buffer, washed and then eluted in a separate tube.

The purified DNA is ready to be used in all demanding molecular biology applications, including restriction digestion, ligation, sequencing, transfection into mammalian cells and in vitro transcription.


TAE or TBE agarose gel (up to 300 mg)Approximately 20 min> 20 µg DNA100 bp – 30 kb60–90%

BR0700301 BR0700302 BR0700303




BR0700401 BR0700402 BR0700403


Solutions, mini filters and VialsSolutions, mini filters and VialsSolutions, mini filters and Vials



42

GenUP™ Plasmid Kit

• Plasmid DNA isolation from Gram-positive and Gram-negative bacteria

APPLICATIONS

FEATURES• Universal kit for both high- copy and low-copy-number plasmid isolation• Fast and simple procedure• High yields of pure plasmid DNA suitable for all applications

DESCRIPTIONbiotechrabbit GenUP Plasmid Kit is designed for fast and efficient plasmid purification from 0.4–10 ml bacterial suspensions in a mini format. After performing an alkaline lysis step and precipitating chromosomal DNA and bacterial proteins, plasmid DNA is bound to a Mini Filter, washed and then eluted in a low-salt buffer. Typically yields are 10–40 µg plasmid DNA from a 5–10 ml bacterial suspension, depending on a plasmid copy number.

The purified plasmid DNA is ready to be used in all demanding molecular biology applications, including enzymatic reactions and sequencing.

Starting material

Extraction timeBinding capacityTypical yield

Average purity

Bacterial cultures: 0.5–5 ml for high-copy-number plasmids, 5–10 ml for low-copy-number plasmidsApproximately 16 minApproximately 40 µg plasmid DNAVariable: 6–20 µg high-copy plasmid DNA is typically purified from 2 ml A260/A280 1.8–2.0

GenUP™ Plasmid Plus Kit

FEATURES• Universal kit for plasmid isolation from Gram-positive and Gram-negative bacteria• Fast and simple procedure• High yields of pure plasmid DNA suitable for all applications

APPLICATIONS• Plasmid DNA isolation from 5–15 ml cultures of Gram-positive and Gram-negative bacteria

DESCRIPTIONbiotechrabbit GenUP Plasmid Plus Kit is designed for fast and efficient plasmid purification from 5.0–15.0 ml bacte-rial suspensions. After performing an alkaline lysis step and precipitating chromosomal DNA as well as bacterial proteins, plasmid DNA is bound to a Mini Filter, washed and then eluted in a low-salt buffer. The purified plasmid DNA is ready to be used in all demanding molecular biology applications, including ligation, sequencing, cloning.

Protocols are available for Gram-negative (such as E. coli) and Gram-positive bacteria.

Starting materialExtraction timeBinding capacityTypical yield

Average purity

Bacterial culture (5–15 ml)Approximately 20 min80 µg plasmid DNA60–70 µg DNA from high-copy plasmid (15 ml culture)A260/A280 1.7–2.0

BR0700201BR0700202 BR0700203




BR0701201BR0701202 BR0701203




40www.biotechrabbit.com

43

GenUP™ Bacteria gDNA Kit

FEATURES• Fast and simple procedure• Genomic DNA from up to 1 × 109 bacteria• High yields of pure DNA for demanding applications

APPLICATIONS• Isolation of bacterial genomic DNA from Gram-positive and Gram-negative bacteria

DESCRIPTIONbiotechrabbit GenUP Bacterial gDNA Kit has been specially developed for quick and easy purification of bacterial genomic DNA from both Gram-negative and difficult to process Gram-positive bacteria.

A combined lysozyme and proteolytic lysis steps allow efficient cell disruption. The DNA is bound to a high-capacity filter, washed and then eluted in a separate tube. The purified DNA is ready to be used in all demanding molecular biology applications, including PCR, enzymatic digestions, cloning and other.

Starting material


1 × 109 Gram-negative or Gram-positive bacterial cells Approximately 45 min> 50 µg DNAVariable; approximately 35 µg

BR0700701BR0700702 BR0700703





GenUP™ Plasmid Midi Kit

• High- or low-copy plasmid isolation DNA from up to 50 ml bacterial cultures

APPLICATIONS

FEATURES• Universal kit for plasmid isolation from Gram-positive and Gram-negative bacteria• Fast and simple procedure• Up to 100 μg of pure plasmid DNA suitable for all applications

DESCRIPTIONbiotechrabbit GenUP Plasmid Midi Kit is designed for fast, simple isolation of plasmid DNA from bacterial cultures of 25 ml for high-copy-number plasmids or up to 50 ml for low-copy-number plasmids. In contrast to anion exchange technology, the plasmid DNA binds to the surface of an optimized GenUP MIDI Filter membrane and is subsequently washed and eluted into a low-salt buffer. The kit allows up to 80 μg plasmid DNA of excellent purity and quality to be recovered without the need for ethanol precipitation. Purified nucleic acids can be used directly for any downstream application..

BR0702001BR0702002 BR0702003

10 preps25 preps 50 preps



Starting material


E. coli overnight culture: 25 ml for high-copy-number plasmids, up to 50 ml for low-copy-number plas-mids and P1 constructsApproximately 45 min100 µg DNAapproximately 50-80 µgA260/A280 1.8–2.0


GenUP™ Mycobacteria gDNA Kit

FEATURES• Fast and simple procedure• gDNA from sputum samples, bronchoalveolar lavage and tissue biopsies• High yields of pure DNA for demanding applications

APPLICATIONS• Isolation of genomic DNA from mycobacteria

DESCRIPTIONbiotechrabbit GenUP Mycobacteria gDNA Kit has been specially developed for quick and easy purification of mycobacterial genomic DNA from various sources including sputum samples, bronchoalveolar lavage and tissue biopsies.

In a first purification step the sample is pre-treated for enrichment of mycobacteria. A following combined lysozyme and proteolytic lysis steps allow efficient cell disruption. The DNA is bound to a high-capacity filter, washed and then eluted in a separate tube. The purified DNA is ready to be used in all demanding molecular biology applications, including PCR, enzymatic digestions, cloning and other.

BR0702201BR0702202 BR0702203




GenUP™ gDNA Kit

• Isolation of genomic DNA from tissues, cells, rodent tail, buccal swaps and paraffin samples

APPLICATIONS

FEATURES• Universal kit for DNA isolation from various eukaryotic starting materials• Fast and simple procedure with >100 µg gDNA filter-binding capacity• High yields of pure DNA

DESCRIPTIONbiotechrabbit GenUP gDNA Kit is designed for a fast and efficient genomic DNA isolation from various sources and different amounts of starting material, such as mammalian tissues (including paraffin-embedded), buccal swabs and eukaryotic cell cultures. After a few initial procedures, the genomic DNA is bound to a Mini Filter, washed and then eluted in a low-salt buffer.

High yields of pure, unshared genomic are ready to be used in all demanding molecular biology applications, including enzymatic reactions and sequencing.

Starting material


Eukaryotic cells (up to 5 × 106), tissue samples (up to 50 mg), rodent tail specimens of 0.5–1 cm in length, paraffin-embedded tissue samples, buccal swabsApproximately 8 min after lysis>100 µg DNAVariable; approximately 65 µgA260/A280 1.7–2.0

BR0700601BR0700602 BR0700603




Starting material

Extraction time

Binding capacityTypical yield

0.2–5 ml sputum sample, up to 1 ml bronchoalveolar lavage or 1–10 mg tissue biopsies (e.g., lymph nodes)Approximately 15 min after the lysis step> 50 μg DNAVariable; yield depends on type of mycobacteria and amount of star-ting material


GenUP™ Blood DNA Kit

FEATURES• Fast and simple procedure• Genomic gDNA from fresh and frozen, EDTA- or citrate-treated blood• Excellent genomic DNA quality in yields of up to 30 µg

APPLICATIONS• Isolation of genomic DNA from up to 400 µl whole blood

DESCRIPTIONbiotechrabbit GenUP Blood DNA Kit is designed for fast isolation of genomic DNA from up to 400 µl whole blood from fresh or frozen samples that have been stabilized with EDTA or citrate. After an efficient lysis step, genomic DNA is bound to a Mini Filter, washed and eluted. The isolation chemistry and extraction protocol are optimized for maximum yield. Including lysis, isolated DNA is available in approximately 24 min. The isolated DNA is suitable for a wide range of different molecular biology applications. Protocols are available for isolating DNA from 200 µl or 400 µl whole blood samples.The GenUP Blood DNA Kit is designed for the use with blood. For other starting material, such as cell-free body fluids (including cerebrospinal fluid, serum, plasma or urine), tissue, stool samples, buffy coat, cultured or isolated cells, swabs, dried blood spots, viruses, fungi, bacteria or parasites, please refer to the GenUP gDNA Kit (cat. no. BR07006), GenUP Bacteria gDNA Kit (cat. no. BR07007), GenUP Plant DNA Kit (cat. no. BR07008) or GenUP Virus DNA/RNA Kit (cat. no. BR07011).

Starting material


Average purity

Fresh or frozen whole blood; stabilized with EDTA or citrate (200 µl or 400µl)Approximately 24 min>60 µg DNAVariable depending on the starting material; approximately 30 µg DNA A260/A280 1.7–2.0

BR0701301BR0701302 BR0701303




GenUP™ Plant DNA Kit

FEATURES• Simple and efficient procedure for plant DNA isolation• Special lysis protocols for different plant materials• High yields of inhibitor-free DNA

APPLICATIONS• Universal kit for isolating genomic DNA from various plant materials

DESCRIPTIONbiotechrabbit GenUP Plant DNA Kit has been specially developed for quick and easy purification of genomic DNA from a wide variety of plant materials, including fresh, frozen or dried samples from leaves, roots, stems and flowers.The kit includes an advanced prefiltration step to remove unlysed tissue. Subsequently, DNA is bound to a Mini Filter and is subsequently washed and eluted in a separate tube. The purified DNA is ready for use in any demanding molecular biology application, including PCR, enzymatic digestions and cloning.This kit provides three buffers for optimized processing with different plant materials. To determine optimal lysis conditions, side-by-side preparation using the three provided protocols are prepared.

BR0700801BR0700802 BR0700803




Starting material


Average purity

Fresh, frozen or dried plant tissue(maximum 100 mg dry weight)Approximately 40 min>50 µg DNAVariable depending on the starting material; approximately 25 µg DNAA260/A280 1.7–2.0


GenUP™ Total RNA Kit

• Universal kit for total RNA isolation from various sources and different amounts of starting material

APPLICATIONS

FEATURES• Fast and simple procedure• High yields of pure RNA• Physical removal of DNA, no DNase treatment, no toxic β-mercaptoethanol

DESCRIPTIONbiotechrabbit GenUP Total RNA Kit has been specially developed for a quick and easy purification of total RNA from eukaryotic cell suspensions, tissues and biopsies, Gram-negative (e.g., E. coli) Gram-positive bacteria and other sources. After few initial procedures, the RNA is bound to a filter, washed and then eluted in a separate tube. DNA is removed physically by binding to a filter wthout any DNase treatment or the use of toxic β-mercaptoethanol. The purified RNA is ready to be used in all demanding molecular biology applications, including cDNA synthesis, northern blot analysis and others.

Starting material


Eukaryotic cells (5 × 106), tissue samples (up to 20 mg), bacterial cells (Gram-positive or Gram-negative, 1 × 109)Approximately 20–40 min100 µg RNAYield is highly dependent on sample type

GenUP™ Micro RNA Kit

FEATURES• Fast and simple procedure• Optimized binding conditions for high yields of small RNAs• Physical removal of DNA, no DNase treatment, no toxic β-mercaptoethanol

APPLICATIONS• Efficient isolation of snRNA, miRNA, siRNA, tRNA, rRNA and mRNA from various starting material

DESCRIPTIONbiotechrabbit™ GenUP Micro RNA Kit is designed for high yields of small RNA molecules and total RNA, includingsnRNA, miRNA, siRNA, tRNA, rRNA and mRNA, without the use of highly toxic β-mercaptoethanol. After usingwell established filter-technology to selectively remove genomic DNA, the RNA is bound, washed and eluted fromthe filter membrane using RNase-free water.

Starting material


Recovery rate

Eukaryotic cells (5 × 106)Fresh or frozen tissue or biopsies (up to 20 mg)Gram-positive and Gram-negative bacteria (up to 1 × 109)Typically 15–40 minutes100 μgDepends on the type and the amount of the starting materialHigh rate of recovery for small RNA molecules

BR0700901BR0700902 BR0700903




BR0701901BR0701902 BR0701903





GenUP™ Blood RNA Kit

• Isolation of up to 8 µg total RNA from 0.5–1.0 ml whole blood

APPLICATIONS

FEATURES• Fast and simple procedure• High-quality RNA isolated from fresh and frozen, EDTA- or citrate-treated blood• Physical removal of DNA, no DNase treatment, no toxic β-mercaptoethanol

DESCRIPTIONbiotechrabbit GenUP Blood RNA Kit is designed for fast isolation of total RNA from up to 1 ml whole blood from fresh or frozen samples that have been stabilized with EDTA or citrate. After an initial lysis step, genomic DNA is bound to a Mini Filter DNA, which can be discarded. RNA is selectively bound to a Mini Filter RNA, washed with two different buffers and eluted. Including lysis, isolated RNA is available in approximately 45 min. The isolated RNA is suitable for a wide range of different molecular biology applications, including RT-PCR.

The GenUP Blood RNA Kit is designed for the use with blood. For other starting material, such as cell-free body fluids (including cerebrospinal fluid, serum, plasma or urine), tissue, stool samples, buffy coat, cultured or isolated cells, swabs, dried blood spots, viruses, fungi, bacteria or parasites, please refer to the GenUP Total RNA Kit (cat. no. BR07009), GenUP Virus RNA Kit (cat. no. BR07010), GenUP Virus DNA/RNA Kit (cat. no. BR07011) or UPzol RNA Isolation Solution (cat. no. BR07001).

Starting material


Fresh or frozen whole blood; stabilized with EDTA or citrate (0.5–1.0 ml)Approximately 45 min>20 µg RNAVariable depending on the starting material; 1–8 µg RNA

GenUP™ Plant RNA Kit

FEATURES• Fast and simple procedure• High-quality RNA isolated from a wide variety of plant samples• Physical removal of DNA, no DNase treatment, no toxic β-mercaptoethanol

APPLICATIONS• Isolation of total RNA from up to 100 mg plant material

DESCRIPTIONbiotechrabbit GenUP Plant RNA Kit has been developed for quick and easy purification of total RNA from plant materials. After initial homogenization and lysis, genomic DNA is bound to a Mini Filter DNA, which can be discarded. RNA is selectively bound to a Mini Filter RNA, washed with two different buffers and eluted. The purified RNA is ready for use in any demanding molecular biology application, including RT-PCR.

Two lysis buffers, Buffer LYSIS R and Buffer LYSIS U, are provided to maximize yield. Most plant material can be pro-cessed with Buffer LYSIS R. In the cases that yield using Buffer LYSIS R is low, use Buffer LYSIS U.

Starting materialExtraction timeBinding capacityTypical yield

Average purity

Plant material (up to 100 mg)30 min after homogenizationApproximately 100 µg RNAVariable depending on the starting material; approximately 70 µg RNAA260/A280 1.7–2.0

BR0701401BR0701402 BR0701403




BR0701501BR0701502 BR0701503





UPzol™ RNA Isolation Solution

• Isolation of RNA using a modified guanidine isothiocyanate/phenol method

APPLICATIONS

FEATURES• Simple and efficient RNA isolation from different sources in scalable volumes• High-quality RNA preparation

DESCRIPTIONbiotechrabbit UPzol RNA Isolation Solution is designed for efficient isolation of total RNA from animal tissues and cells, bacterial cells, plants and other material in variable amounts. The extraction method is based on a time-saving, one-step liquid phase separation.The UPzol RNA Isolation Solution is a mono-phase solution containing phenol and guanidine thiocyanate. After the addition of chloroform and subsequent centrifugation, the homogenate is separated into three phases:• Colorless aqueous phase containing RNA (upper)• White interphase (middle)• Colored organic phase (lower)

RNA is precipitated from the upper aqueous phase using alcohol. The UPzol RNA Isolation Solution provides high-quality and high-integrity RNA, which can be used for all downstream applications, including northern analysis, cDNA synthesis, RT-PCR, dot-blot hybridization, poly A+ selection, in vitro translation, cloning and RNase assays.

Starting material

Extraction timeTypical yield

Tissue (100 mg), monolayer cells (3 cm dish), suspension of animal, plant, yeast or bacterial cells (5 × 106)Approximately 1 hDepends on the type and amount of starting material

GenUP™ Virus DNA/RNA Kit

FEATURES• Universal kit for simultaneous RNA and DNA isolation from different starting materials• Fast and simple procedure, sample specific protocols• High yields of pure RNA and DNA • Physical removal of DNA, no DNase treatment, no toxic β-mercaptoethanol

APPLICATIONS• Simultaneous viral DNA and RNA isolation from various sources• Excellent performance for unknown viruses

DESCRIPTIONbiotechrabbit GenUP Virus DNA/RNA Kit has been specially developed for quick and easy isolation of viral RNA and DNA. The kit is especially useful when the origin of the virus is unknown. Viral double-stranded DNA and single-stranded RNA are simultaneously isolated from eukaryotic samples, including plasma, serum, and other body fluids as well as cell cultures, tissues, and buccal swabs.The unique binding membrane of our high-capacity Mini Filters guaranties high yields. A high concentration of purified nucleic acid can be achieved with flexible elution volumes. The kit includes carrier RNA (CARRIER).After a few initial procedures, the viral nucleic acids are bound to a Mini Filter, washed and then eluted in a separate tube. The purified nucleic acids are ready to be used in all demanding molecular biology applications, including cDNA synthesis, northern blot analysis, qPCR and RT-PCR.

Starting material


Eukaryotic cells (up to 5 × 106), serum, plasma, cell-free body fluids, cell culture supernatants (150 µl), tissue samples, biopsies (up to 20 mg), paraffin-embedded tissues, buccal swabsApproximately 25 minYield is highly dependent on sample type

BR0700101BR0700102

100 preps 200 preps

100 ml UPzol RNA Isolation Solution2 × 100 ml UPzol RNA Isolation Solution


BR0701101BR0701102 BR0701103





GenUP™ Virus RNA Kit

• Virus RNA isolation from plasma, serum, urine and other body fluids, cell cultures, tissues, and buccal swabs

APPLICATIONS

FEATURES• Universal kit for isolating viral RNA from various starting materials• Fast and simple procedure, flexible elution volumes• High yields of pure RNA • Physical removal of DNA, no DNase treatment, no toxic β-mercaptoethanol

DESCRIPTIONbiotechrabbit GenUP Virus RNA Kit has been specially developed for quick and easy isolation of viral RNA. Viral single-stranded RNA can be isolated from eukaryotic samples including plasma, serum, urine, and other body fluids as well as cell cultures, tissues, and buccal swabs.

The unique binding membrane of our high-capacity Mini Filters guaranties high yields. A high concentration of purified RNA can be achieved with flexible elution volumes. The kit includes carrier RNA (CARRIER).

After few initial procedures, the viral RNA is bound to a Mini Filter, washed and then eluted in a separate tube. The purified RNA is ready to be used in all demanding molecular biology applications, including cDNA synthesis, northern blot analysis, qPCR and RT-PCR.

Starting material


Eukaryotic cells (up to 5 × 106), serum, plasma, cell-free body fluids, cell culture supernatants (150 µl), tissue samples, biopsies (up to 20 mg), paraffin-embedded tissues, buccal swabsApproximately 25 minYield is highly dependent on sample type

BR0701001BR0701002 BR0701003




GenUP™ FFPE Paraffin Removal Solution

• Removal of paraffin from formalin-fixed, paraffin-embedded (FFPE) sample slices

APPLICATIONS

FEATURES• One-step deparaffinization of FFPE slices• No toxic or flammable chemicals

DESCRIPTIONbiotechrabbit GenUP FFPE Paraffin Removal Solution is an odor-free, biofriendly and innovative solution to deparaffinize FFPE slices within minutes in a single step. This product replaces conventional procedures that involve toxic and/or flammable solvents. After removal of paraffin from FFPE slices with GenUP FFPE Paraffin Removal Solution, continue with the genomic DNA purification kit of your choice.

BR0701601

BR0701602

10 ml

50 ml

10 ml GenUP FFPE Paraffin Removal Solution5 × 10 ml GenUP FFPE Paraffin Removal Solution



GenUP™ Bisulfite Kit

FEATURES• Fast and easy bisulfite conversion with high conversion rates• Flexible use with a wide range of sample types• Including subsequent desulfonation and DNA cleanup

APPLICATIONS• Bisulfite conversion of unmethylated cytosines to uracils for epigenomics studies

DESCRIPTIONbiotechrabbit GenUP Bisulfite Kit provides a fast and easy method for the conversion of unmethylatedcytosines to uracils enabling DNA methylation studies. The kit is designed for high conversion rates, extremely easy application and exceptionally fast, reliable results. Denaturation and bisulfite treatment are performed in a single tube in approximately 3 h. After conversion, the DNA is cleaned up and desulfonated on a Mini Filter and can be used for downstream applications, such as PCR and sequencing.

Starting materialExtraction time

Purified DNA (1 pg to 10 µg)Conversion reaction: 45 minPurification and desulfonation: 45 min

BR0701701BR0701702 BR0701703




GenUP™ Bisulfite Complete Kit

FEATURES• Fast and easy bisulfite conversion with high conversion rates• Flexible use with a wide range of sample types including cells, tissues and FFPE samples• Including subsequent desulfonation and DNA cleanup• Isolation of both modified and unmodified genomic DNA in parallel

APPLICATIONS• Isolation of genomic DNA with subsequent bisulfite conversion of unmethylated cytosines to uracils and subsequent desulfonation and DNA cleanup.

DESCRIPTIONbiotechrabbit™GenUP Bisulfite Kit provides a fast and easy method for the isolation of genomic DNA from various sources in combination with conversion of unmethylated cytosines to uracils enabling DNA methylation studies (see Fig. 1 and Fig. 2). The kit is designed for high conversion rates, extremely easy application and exceptionally fast, reliable results. Both modified and unmodified genomic DNA can be isolated in parallel in one experiment setup. Denaturation and bisulfite treatment are performed in a single tube in approximately 3 h. After conversion, the DNA is cleaned up and desulfonated on a Mini Filter and can be used for downstream applications, such as PCR and sequencing. The purified DNA can then be stored at −20°C or used for subsequent applications.

BR0702101BR0702102 BR0702103




Purified DNA (1 pg to 10 μg), cells (up to 5 × 105), fresh tissue (max. 1 mg),FFPE tissue sections (1–3 sections max. 10 μm each), FFPE tissue punch biopsy core (max. 10 mg), bronchial aspirates, swabs, perito-neal cavity fluid, pleural effusions, sputum, urine sedimentLysis approx. 20 min to 48 h, depen-ding on the sample type, conversion reaction 45 min, purification and desulfonation 45 min

Starting material

Extraction time


Electrophoresis

Stable and ready-to-use, biotechrabbit DNA and protein ladders provide dyes, allowing the progress of gel elec-trophoresis to be monitored. The location and intensity of ladder bands in gels can be used to estimate the size and, for DNA, quantity of other bands in the gel. Nuclease-free DNA Loading Dye is also available.

• Time saving — ready to use for convenience

• Reliable — high purity and stability

DNA Electrophoresis Ladders and Loading Dye

1 kb DNA Ladder with 6× Loading DyeBR0800101 100 lanes 500 µl 1 kb DNA Ladder

1 ml DNA Loading Dye

Package contentCat. no. Size• DNA sizing and approximate quantification on agarose gels

APPLICATIONS

FEATURES• Ready to use DNA ladders ideal for DNA sizing and gel quantification• Pure and stable — retain sharp bands after 6 months storage at room temperature• Supplied with 6× Loading Dye for sample DNA

DESCRIPTIONbiotechrabbit DNA electrophoresis ladders are mixtures of exceptionally purified DNA fragments created either by PCR or by digesting proprietary plasmids with restriction enzymes. Ladders are ready to use and suitable not only for DNA sizing but also for approximate DNA quantification in gels. For convenience, ladders have increased intensity reference bands and indicated DNA amount in nanograms for every band.

Every ready-to-use ladder is supplied with the nuclease-free Loading Dye Solution, which ensures optimal migration and quantification of your DNA probes. It includes three electrophoresis tracking dyes (xylene cyanol, bromophenol blue and orange G), allowing the process of the DNA through the gel to be visualized.

100 bp DNA Ladder with 6× Loading DyeBR0800201 100 lanes 500 µl 100 bp DNA Ladder

1 ml DNA Loading Dye50 bp DNA Ladder with 6× Loading DyeBR0800401 100 lanes 500 µl 50 bp DNA Ladder

1 ml DNA Loading DyeDNA Loading Dye, 6×BR0800301 5 ml 5 × 1 ml DNA Loading Dye

Ready-to- use ladder:Range | bands:Reference:

1 kb DNA Ladder250–10,000 bp | 131000 and 3000 bp

100 bp DNA Ladder100–3000 bp | 12500 and 1500 bp

50 bp DNA Ladder50–1500 bp | 17200 and 500 bp

DNA Loading Dye, 6×Migration of dyes in 1% TAE agarose gel

Xylene cynol FF ~4 kb

Bromophenol blue ~400 bp

Orange G ~50 bp

TriColor Broad Protein Ladder (3.5–245 kDa)BR0900101 100 minigel

appl. 500 µl TriColor Broad Protein Ladder (3.5–245 kDa)

Package contentCat. no. Size• Approximate protein sizing on SDS PAGE and western blots• Monitoring protein gel electrophoresis and western transfers

APPLICATIONS

FEATURES• Ready-to-use prestained protein ladders covering 3.5–245 kDa and 10–180 kDa ranges• Pure and stable — retain sharp bands after 3 months storage at 4°C• Protein bands in three colors with easily recognizable colored reference bands

DESCRIPTIONbiotechrabbit TriColor Protein Ladders facilitate approximate molecular-weight estimation of proteins on denaturing polyacrylamide gels, monitoring protein separation during electrophoresis and verification of western transfer efficiency to membranes (polyvinylidene difluoride, nylon or nitrocellulose). The ladders provide three colored reference bands.

TriColor Broad Protein Ladder is ready for immediate use and is notably stable: sharp bands are produced after storage for 2 weeks at room temperature or 3 months at 4°C, eliminating the need to thaw before loading.

Protein Electrophoresis Ladders

TriColor Protein Ladder (10–180 kDa)BR0900201 100 minigel

appl. 500 µl TriColor Broad Protein Ladder (10–180 kDa)

Ready-to- use ladder:Range | bands:Reference:

TriColor Protein Ladder (10–180 kDa)10–180 kDa | 1025 and 75 kDa

TriColor Broad Protein Ladder (3.5–245 kDa)3.5-245 kDa | 13

25 and 75 kDa

Migration patterns in different electrophoresis conditions. The apparent molecular weight of each protein (kDa) was determined by calibrating against unstained proteins under same conditions.


Enzymes for Molecular Biology

biotechrabbit enzymes are suitable for the most demanding molecular biology applications. Selected for specific characteristics and excellent performance, our enzymes provided in exceptionally pure preparations to ensure the integrity of your reactions.

• High performance — success in your

molecular biology applications

• Exceptional purity — reliable results in

demanding reactions

Product Applications Features

phi29 DNA Polymerase

StranDisplace™ Thermostable DNA Polymerase

T4 DNA Ligase, Rapid

Shrimp Alkaline Phosphatase

Heat labile Uracil–DNA Glycosylase

Uracil–DNA Glycosylase

Heat labile dsDNase

Proteinase K

T7 RNA Polymerase

Rolling circle amplification, multiple displacement amplification, whole genome amplification, protein-primed DNA ampli-fication, RNA-primed DNA amplification

Loop-mediated isothermal amplification, sequencing

Blunt and sticky end DNA ligation for cloning, joining linkers or adaptors to double-stranded DNA, self-circularization of linear DNA

Removal of nucleotides from PCR prior to sequencing, dephosphorylation of restriction-digested vectors to prevent religation prior to cloning, rapid dephosphorylation of RNA, proteins or other biomolecules

Elimination of carry-over contamination in PCR, RT-PCR, qPCR and RT-qPCR, post-PCR analysis such as cloning and sequencing, analysis of ancient DNA

PCR carry-over contamination control

Integration into cDNA synthesis protocols, removal of genomic DNA from RNA preparations, removal of contaminating DNA from PCR, qPCR and RT-qPCR, purification of DNA polymerases

Nonspecific protein degradation, nucleic acid purification, deactivation of nucleases in enzymatic reactions

Generation of mRNA for in-vitro translation,RNA labeling or gene silencing

Huge DNA yields, accurate amplification, highest processivity with exceptionally strong strand-displacement activity

Exceptional purity for demanding applications, strong strand-displacement activity

Exceptional purity and high concentration for demanding applications, rapid ligation, supplied with two buffers for standard and fast ligation protocols

Completely inactivated after 5 min at 65°C, active in com-mon restriction enzyme and PCR buffers; no need for extra addition of buffer of ions, exhibits excellent stability at 4°C and room temperature

Completely and irreversibly heat inactivated without addition of agents or inhibitors, active in common PCR and RT-PCR buffers

High quality for a fair price

Heat-inactivated at high temperatures, degrades genomic DNA in RNA preps, does not affect RNA quality or quantity

Exceptional purity suitable for the most sensitive applications

High-yield in vitro transcription with strictly promotor-specific activity


phi29 DNA Polymerase, 10 U/µl

• Rolling circle amplification• Multiple displacement amplification• Whole genome amplification• Protein-primed and RNA-primed DNA amplification

APPLICATIONS

DESCRIPTIONbiotechrabbit phi29 DNA Polymerase is an exceptionally pure DNA polymerase for demanding applications. The enzyme is purified from a recombinant E. coli strain carrying the phi29 DNA Polymerase gene from bacteriophage phi29.

The enzyme is a highly processive DNA polymerase (up to 70,000 base insertions per binding event) with a powerful strand-displacement activity and a 3'→ 5' proofreading exonuclease function.

phi29 DNA Polymerase proofreading activity acts preferentially on single-stranded DNA or RNA. Therefore, to avoid primer degradation during the DNA synthesis, 3'modified (protected) primers are highly recommended.

StranDisplace™ Thermostable DNA Polymerase, 8 U/µl

FEATURES• Exceptionally pure thermstable DNA polymerase for demanding applications• Strong strand-displacement activity

APPLICATIONS• Loop-mediated isothermal amplification• Sequencing

DESCRIPTIONbiotechrabbit StranDisplace Thermostable DNA Polymerase is an exceptionally pure DNA polymerase for applications in which strong strand-displacement activity at elevated temperatures is required.

The StranDisplace Thermostable DNA Polymerase is a thermophilic DNA polymerase with a strong strand-displacement activity and is deficient in both proofreading (3'→ 5') and nick-translation (5'→ 3') nuclease activities. The polymerase can be used for the same applications as Bst DNA Polymerase Large Fragment and Bsm DNA Polymerase.

FEATURES• Huge DNA yields, accurate amplification• Highest processivity DNA polymerase with exceptionally strong strand displacement activity• Exceptionally pure phi29 DNA Polymerase for demanding applications

BR1100101

BR1100102

250 U

1000 U

25 µl phi29 DNA Polymerase 250 µl 10× phi29 Reaction Buffer100 µl phi29 DNA Polymerase 1 ml 10× phi29 Reaction Buffer


BR1100201 8000 U 1 ml StranDisplace Thermostable DNA Polymerase4 × 1.5 ml 10× SD Reaction Buffer



T4 DNA Ligase, Rapid, 600 U/µl

• Blunt and sticky end DNA ligation for cloning• Joining of linkers or adaptors to double-stranded DNA• Self-circularization of linear DNA

APPLICATIONS

FEATURES• Exceptionally pure high-concentration T4 DNA Ligase for demanding applications• Rapid ligation• Supplied with two buffers for fast and usual ligation protocols

DESCRIPTIONbiotechrabbit T4 DNA Ligase, Rapid, is an exceptionally pure, highly concentrated ligase for applications in which high enzyme concentrations are required. It is especially recommended for fast ligations.

T4 DNA Ligase, Rapid, is supplied with two buffers: the common 10× Ligation Buffer for typical 1 h or overnight ligation and the 2× Rapid Ligation Buffer containing PEG for fast 5–10 minutes ligation or ligation of low-concentration or blunt-end DNA.

T4 DNA Ligase catalyzes the formation of a phospho-diester bond between the terminal 5' phosphate and the 3' hydroxyl groups of duplex DNA or RNA. The enzyme efficiently joins blunt and cohesive ends and repairs single stranded nicks in duplex DNA, RNA or DNA–RNA hybrids.

BR1100301

BR1100302

60000 U

180000U

100 µl T4 DNA Ligase, Rapid1.5 ml 2× Rapid Ligation Buffer500 µl 10× Ligation Buffer3 × 100 µl T4 DNA Ligase, Rapid3 × 1.5 ml 2× Rapid Ligation Buffer3 × 500 µl 10× Ligation Buffer


55

Shrimp Alkaline Phosphatase, 1 U/µl

FEATURES• Quickly and easily removes 5' phosphates from DNA, RNA, dNTPs and proteins• Completely inactivated after 5 min at 65°C• Active in common restriction enzyme and PCR buffers• Exhibits excellent stability at 4°C and room temperature

APPLICATIONS• Removal of nucleotides from PCR prior to sequencing• Dephosphorylation of restriction-digested vectors to prevent religation prior to cloning• Rapid dephosphorylation of RNA, proteins or other biomolecules

DESCRIPTIONbiotechrabbit Shrimp Alkaline Phosphatase is a multipur-pose alkaline phosphatase that dephosphorylates most biomolecules and is fully inactivated by heating for 5 min at 65°C. The enzyme is stable at room temperature and active in common buffers for restriction digestion and PCR.

Dephosphorylation of restriction-digested plasmid to prevent religation prior to cloning Shrimp Alkaline Phosphatase provides an efficient method to reduce the religation of vectors during cloning of DNA fragments by dephosphorylating restriction-digested plasmids. The enzyme dephosphorylates all types of DNA termini — 3'-protruding, blunt and 5'-protruding — and is either added to the restriction mixture or after digestion. Subsequently, Shrimp Alkaline Phosphatase is efficiently heat-inactivated. During the ligation of plasmid with DNA fragments, the background of unwanted “empty” clones is reduced to less than 5%.

PCR cleanup prior to sequencing The combination of Exonuclease 1 (not included) and Shrimp Alkaline Phosphatase provides a simple and fast method for PCR cleanup. Both enzymes are simply added to the PCR mixture, incubated and heat-inactivated. Shrimp Alkaline Phosphatase dephosphorylates nucleotides and primers, while Exonuclease 1 degrades primers, leaving a PCR product that can be directly used for sequencing or genotyping.

BR1100601BR1100602

1000 U5000 U

1 ml Shrimp Alkaline Phosphatase5 × 1 ml Shrimp Alkaline Phosphatase



Heat Labile Uracil–DNA Glycosylase, 1 U/µl

FEATURES• The only Uracil–DNA glycosylase that is completely and irreversibly heat inactivated• Heat-labile without any addition of agents or inhibitors• Active in common PCR and RT-PCR buffers

APPLICATIONS• Eliminates carry-over contamination in PCR, RT-PCR, qPCR and RT-qPCR• Enables downstream post-PCR analysis such as cloning and sequencing• Analysis of ancient DNA

DESCRIPTIONbiotechrabbit Heat Labile Uracil–DNA Glycosylase selectively degrades uracil-containing PCR products. After performing PCR or RT-PCR using dUTP instead of dTTP, PCR products remain intact after treatment with Heat Labile Uracil–DNA Glycosylase, whereas contaminating DNA (i.e., not amplified) is degraded. Heat Labile Uracil–DNA Glycosylase is completely and irreversibly inactivated by moderate heat treatment at 50⁰C, allowing contamination control in RT-qPCR. The enzyme hydrolyses the N-glycosylic bond between the deoxyribose sugar and the base in uracil-containing DNA leaving an abasic (apyrimidinic) site in DNA but does not modify uracils in RNA.

Heat Labile Uracil–DNA Glycosylase is highly active at 20–40°C. No cofactors or divalent cations are required for activity, and the enzyme is active in most PCR and RT-PCR buffers. Although the enzyme is active a pH 6.5–9.0, the optimal pH 7.5 is in 50 mM NaCl.

BR1100701 BR1100702

100 U

1000 U

100 µl Heat Labile Uracil-DNA Glycosylase1 ml Heat Labile Uracil-DNA Glycosylase


FEATURES• Exceptionally pure Uracil–DNA Glycosylase for demanding applications• High value for a fair price

APPLICATIONS• PCR carry-over contamination control• Site-directed mutagenesis• Glycosylase-mediated single-nucleotide polymorphism detecion

DESCRIPTIONbiotechrabbit Uracil–DNA Glycosylase (UDG) is an exceptionally pure enzyme that is especially useful for PCR carry-over contamination control. Amplification of contaminating templates remaining in the work environment from previous experiments is one of the most common PCR contamination problems. The use of Uracil–DNA Glycosylase (UDG) and dUTP (dNTP/dUTP Mix) helps prevent carry-over contamination.The enzyme shows no measurable activity with short oligonucleotides (<6 bases) and RNA substrates. Uracil–DNA Glycosylase is purified from a recombinant E. coli strain carrying the Uracil–DNA glycosylase gene from E. coli K-12.Uracil–DNA Glycosylase degrades DNA containing uracil. To prevent carry-over contamination, use dUTP or dTTP/dUTP rather than dTTP alone for PCR. After PCR with dUTP, the PCR products are substrates for UDG and any DNA containing uracil is degraded during the brief incu-bation with Uracil–DNA Glycosylase. This step eliminates contaminating DNA from previous experiments.

BR1100401

BR1100402

200 U

1000 U

100 μl Uracil–DNA Glycosylase300 μ 10x UDG Reaction Buffer 500 μl Uracil-DNA Glycosylase1.5 ml 10x UDG Reaction Buffer


Uracil–DNA Glycosylase, 2 U/µl


57

Heat Labile dsDNase, 2 U/µl

FEATURES• Provides endonuclease activity specific for double-stranded DNA • Is easily heat-inactivated at low temperatures• Degrades genomic DNA in RNA preps• Does not affect RNA quality or quantity

APPLICATIONS• Easy integration into cDNA synthesis protocols• Removal of genomic DNA from RNA preparations• Removal of contaminating DNA from PCR, qPCR and RT-qPCR• Purification of DNA polymerases

DESCRIPTIONbiotechrabbit Heat Labile dsDNase is an endonuclease that cleaves phosphodiester bonds in DNA to yield oligo-nucleotides with 5'-phosphate and 3'-hydroxyl termini. The enzyme has a particularly strong preference for double-stranded DNA (dsDNA), thus double-stranded DNA is degraded, leaving single-stranded DNA and RNA intact. The enzyme is completely and irreversibly inactivated with 5 min at 55°C in the presence of 1 mM DTT and pH 8.0. Heat Labile dsDNase is highly active at 20–40°C .

BR1100801BR1100802

250 U1000 U

125 µl Heat Labile dsDNase4 × 125 µl Heat Labile dsDNase


Proteinase K

FEATURES• Exceptional purity suitable for the most sensitive applications• PCR-grade for highest performance• Stable lyophilizate

APPLICATIONS• Nonspecific protein degradation• Nucleic acid purification• Deactivation of nucleases in enzymatic reactions

DESCRIPTIONbiotechrabbit Proteinase K is an active endopeptidase that is effective with native proteins, allowing endogenous RNases and DNases to be inactivated rapidly. The robust enzyme is stable over a wide pH range (4–12.5) and remains fully active for several hours when incubated at pH 6.5–9.5. The exceptional purity of the enzyme ensures that it is ideally suited for preparing PCR templates, as it is free of RNases, DNases and DNA.

BR1100901 BR1100902

30 mg 150 mg

30 mg Proteinase K lyophilized 5 × 30 mg Proteinase Klyophilized




T7 RNA Polymerase, 50 U/µl

FEATURES• High-yield in vitro transcription• Strictly promoter-specific activity• No promoter cross-talk• Free from endonuclease, exonuclease, protease, RNase

APPLICATIONS

• Preparation of homogeneously labeled RNA probes for hybridization (e.g., radioactive, biotin, DIG-11-UTP)• Generation of mRNA for in vitro translation, gene expression or splicing studies• Production of template for cDNA synthesis• In vitro synthesis of capped RNA transcripts• Synthesis of antisense RNA and dsRNA for RNA interference or gene silencing

DESCRIPTIONT7 RNA Polymerase is an extremely promoter-specific, DNA-dependent polymerase from the T7 bacteriophage. The enzyme requires Mg2+ ions and the presence of the T7 promoter in the template DNA to synthesize RNA in the 5́ → 3´ direction.The combination of the high-specificity binding, which reduces cross-talk with other promoters, and the rigorous purity testing, which confirms absence of other enzymatic activities, ensures that the T7 RNA Polymerase transcri-bes large amounts of RNA from sequences downstream of the T7 promoter. By using T7 promoters in opposite orientation, double-stranded RNA can be generated. The synthesized RNA is suitable for use in a wide range of applications, including hybridization (e.g., blot, in situ, and microarray), RNA protection, capped RNA and antisense RNA.

BR1101201 BR1101202

5000 U

25000 U

100 μl T7 RNA Polymerase500 µl 10× Reaction Buffer500 μl T7 RNA Polymerase2 × 1.5 ml 10× Reaction Buffer


NOTES



Cell-Free Protein Synthesis

Cell-free protein synthesis overcomes the limitations of traditional cell-based methods. Advantages are ease of use, speed, throughput and the flexible character of the system. Expression conditions can be controlled and easily adapted by adding supplements. It is possible to express toxic proteins and to incorporate labeled amino acids into the protein of interest.Laborious up- and downstream procedures of traditional in vivo methods are avoided as this technique does not requi-re gene transfection or cell culture. By using linear PCR ba-sed templates to express the gene of interest, time-consu-ming cloning procedures are completely avoided.biotechrabbit expression systems are based on extracts from E. coli, wheat germ and insect cells. All components are optimized for highest productivity.

• From gene to protein within one day

• No transfection, no cell culture

• Expression of toxic proteins

• Easy site-specific labeling of proteins

• Excellent reproducibility

• High-throughput screening approaches

• Dedicated His-tag protein purification kits

Cell-Free Expression Workflow

Continuous Exchange Cell-Free (CECF) Technology

Batch CECFCell-free protein expression in batch format is highly suitable for screening applications because of the fast and easy setup. However, due to inhibition from byproducts and depletion of subtrates, yields using this process are limited.Highest yields are achieved with continuous exchange during cell-free expression (CECF), which allows pro-tein expression to continue for 24 hours. CECF relies on diffusion through a semipermeable membrane between the reaction and the feeding chamber. The reaction chamber contains the machinery for protein expression, while the feeding chamber continuously supplies amino acids, NTPs and energy-regenerating compounds. During the reaction, inhibitory byproducts diffuse out off the reaction chamber.In combination with the optimized high-yield (HY) lysates, synthesis rates using CECF are much higher compared to conventional cell-free expression systems.

In vitro transcription and translation

Cell-free Protein Synthesis S. 60

GENE OF INTEREST

GENE OF INTEREST

P E O

N F R

I

E T T

N

O S

R E T

I

T A G

Active components

Protein product Semipermeable membrane

Gene template

Cell-free lysate

+

Incubation

Protein analysis or functional assay

Gene template

Transcription

T7P RBS Tag T7T

Translation Lysate

T7 RNAPolymerase

mRNA

Protein

Substrates

Inhibitory byproducts


Cell-Free Synthesis Kits

biotechrabbit provides cell-free expression kits for the syn-thesis of a broad range of proteins for screening, characte-rization studies, functional assays or structural analysis.The platform uses cell lysates of different origin and it enab-les scalable in vitro protein expression with unique patented features to generate exceptional high protein yields of up to 6 mg/ml.Dedicated kits for the expression of functional proteins containing disulfide bonds or post translational modifica-tions are available as well as various supplementary kits for gene template generation, solubility enhancement, additional amino acids or chaperones. Starting from a DNA template, the system enables protein expression with yields ranging from micrograms to tens of milligrams. Start expression from linear or circular template according to your needs.

• Convenient workflow, all components included

• Expression of different protein classes

• Continuous exchange during cell-free

expression (CECF) for yields of up to 6 mg/ml

• Incorporation of labeled amino acids

• Enhanced high-yield (HY) buffer compositions

• Compatible with T7 expression vectors

Cell-free synthesis kit yield sizeTime(hrs)

Template(No. see p. 70) Mode Proteins expressed

RTS 100 E. coli HY Kit

RTS 500 ProteoMasterE. coli HY Kit


EasyXpress E. coli Kit

RTS 100 E. coli Fab Kit

RTS 100/500 E. coliDisulfide Kits

RTS 100 Wheat Germ Kit

RTS 100/500 Wheat Germ CECF Kit

RTS 100 Insect Disulfide Kit

RTS 100 Insect Membrane Kit

RTS 100 Site-Specific Label Kit

Reaction

20 µg

6 mg

50 mg

20 µg400 µg

30 µg

80 µg2.5 mg

5 µg

50 µg1 mg

2 µg

2 µg

150 µg

50 µg

1 ml

10 ml

50 µl1 ml

400 µl

50 µl1 ml

50 µl

50 µl1 ml

100 µl

50 µl

50 µl

1–4

6–24

6–24

1

6–24

6–24

3

6–24

4

4

1–24

Linear: 1, 4, 5Vector: 7, 9

Vector: 7, 9

Vector: 7, 9

Linear: 3Vector: 10

Linear: 1, 4Vector: 7, 9

Vector: 7, 9

Linear: 2Vector: 8

Linear: 2Vector: 8

Linear: 1, 3, 4Vector: 6, 7, 9

Linear: 1, 3, 4, 5Vector: 6, 7, 9

Linear: 1, 4, 5Vector: 7, 9

Batch

CECF

CECF

Batch

Batch

CECF

Batch

CECF

Batch

Batch

CECF

*

*

*

*

*

*

*

( )

Standard soluble

Complex and large

Disulfide-bonded

Fab antibodies

Glycosylated

Membrane

RTS: Rapid translation system Linear: PCR-generated linear expression templates Vector: expression plasmid * using detergents



FEATURES• Compatible with linear templates for fast screening without cloning• Convenient reaction setup• Improved high yield biochemistry for up to 20 μg protein per reaction

APPLICATIONS• Fast parallel microgram-scale protein expression

DESCRIPTIONThe biotechrabbit RTS 100 E. coli HY Kit contains a modified high-yield E. coli lysate to drive coupled in vitro transcription and translation reactions. The enhanced kit can express up to 20 µg protein (50 µl reaction) in 4 hours using a batch mode. The RTS 100 E. coli HY kit is compa-tible with both linear templates and plasmids, making it the product of choice for protein-expression screening and optimization.Each kit contains everything you need to perform protein-expression in the delivered microplate or, alternatively, in microtubes.

Proteins expressed

Reaction time / yield Reaction modeTemplate

Standard soluble proteinsMembrane proteins1–4 h / 20 µgBatchRTS Linear Template Kit PlusRTS Linear Template Fab KitRTS 100 E. coli LinTempGen Set, HisRTS pIX3.0 VectorRTS pIVEX E. coli His-tag, 2nd Gen. V.

BR1400101

BR1400102

24 rxn of 50 µl96 rxn of 50 µl

Reagents for coupled transcription/translation reactionsReagents for coupled transcription/translation reactions


RTS 500 ProteoMaster E. coli HY Kit

FEATURES• Unique CECF technology to ensure high yields• Enable expression of toxic proteins that cannot be produced in vivo• Incorporation of labeled methionine into synthesized proteins

APPLICATIONS• Optimized medium-scale, cell-free expression of up to 6 mg protein• Protein synthesis for NMR spectroscopy or X-ray crystallography

DESCRIPTIONThe biotechrabbit RTS 500 ProteoMaster E. coli HY Kit is designed and optimized for the expression of up to 6 mg of protein from plasmid templates in highly efficient coupled transcription/translation reactions. This is achieved by utilizing a biochemically enhanced high-yield (HY) E. coli lysate and the RTS 500 CECF reaction device.

Due to preparative expression yields, the RTS 500 Proteo-Master E. coli HY Kit is suited for the production of protein for NMR spectroscopy or X-ray crystallography. The kit design allows to use amino-acid mixtures of your choice. Methionine (included in the kit separately from other amino acids) can be easily substituted by sele-no-methionine (not included). Other amino acids can be exchanged by preparing tailored amino-acid mixtures using the RTS Amino Acid Sampler.

The RTS 500 ProteoMaster E. coli HY Kit is optimized for use in a thermomixer. The continuous agitation of the device guarantees optimal support of the CECF process, which is required for efficient protein expression.

Proteins expressed


Standard soluble proteinsMembrane proteins6–24 h / 6 mgCECFRTS pIX3.0 VectorRTS pIVEX E. coli His-tag, 2nd Gen. V.

BR1400201 5 rxn of 1 ml

Reagents for coupled transcription/translation reactions



FEATURES• Unique CECF technology to ensure high yields• Expression of toxic proteins that cannot be produced in vivo• Incorporation of labeled methionine into proteins

APPLICATIONS• Large-scale, cell-free expression of up to 50 mg protein

DESCRIPTIONOne reaction of the biotechrabbit RTS 9000 E. coli HY Kit yields ten times more total protein (up to 50 mg) than an RTS 500 reaction. The kit provides a reaction volume ten times higher than the RTS 500 E. coli HY Kit, while main-taining a similar productivity per milliliter of lysate.The RTS 9000 Reaction Device is composed of a reaction compartment and a feeding compartment. The 10 ml reaction compartment, where the coupled transcription/translation reactions take place, is separated from the 100 ml feeding compartment by a semipermeable membrane (cut-off 10 kDa).

Proteins expressed


Standard soluble proteinsMembrane proteins6–24 h / 50 mgCECFRTS pIX3.0 VectorRTS pIVEX E. coli His-tag, 2nd Gen. V.

BR1400301 1 rxn of 10 ml




EasyXpress E. coli Kit

FEATURES• Fast procedure — from gene to protein in a single day• High yields — up to 1 mg protein per ml reaction volume• Convenience — color-coded reagents and easy-to- follow protocols

APPLICATIONS• For fast in vitro synthesis of recombinant proteins with high yields

DESCRIPTIONThe biotechrabbit EasyXpress E. coli Kits are designed for rapid cell-free synthesis of recombinant proteins. The kits use highly productive cell-free E. coli lysates, which cont-ain all transcription and translation components required for efficient protein synthesis at any scale. Multiple protein constructs can be screened in small scale reactions.The E. coli EasyXpress Kits enable synthesis of active, full-length recombinant proteins directly from plasmid DNA or PCR product templates. In combination with the protein synthesis procedure, all steps from template generation to protein purification can be performed in a single day.The kits express genes using linear templates. Multiple expression constructs may easily be generated by PCR to be used for protein synthesis. Linear template can be cloned into the compatible cloning vector pIX3.0 Vector to upscale using RTS E. coli kits.

Proteins expressed


Standard soluble proteinsMembrane proteins1 h / 20 µg (50 µl), 400 µg (1 ml)BatchEasyXpress Linear Template Kit PlusEasyXpress pIX3.0 Vector

BR1402001

BR1402002

BR1402003

5 rxn of 50 µl 20 rxn of 50 µl 4 rxn of 1 ml

Reagents for coupled transcription/translation reactions Reagents for coupled transcription/translation reactions Reagents for coupled transcription/translation reactions



RTS 100 E. coli Fab Kit

FEATURES• Accelerated microgram-scale protein expression• Compatible with PCR-generated templates and plasmids

APPLICATIONS• Expression of Fab antibody fragments and disulfide proteins for rapid functional testing• Generation of intra- and intermolecular disulfide bonds

DESCRIPTIONThe biotechrabbit RTS 100 E. coli Fab Kit is designed for rapid functional screening of antibody fragments and disulfide proteins and facilitating the assignment of biolo-gical functions to genes. Each reaction produces enough protein for small-scale affinity purification.The RTS 100 E. coli Fab Kit contains a modified E. coli lysa-te that is enriched in chaperones and disulfide isomerase activity and maintains a defined redox potential to drive coupled in vitro transcription and translation reactions for the synthesis of proteins with intra- and intermolecular disulfide bonds. The kit is compatible with both linear tem-plates and plasmids, making it the product of choice for functional screening of disulfide proteins and optimization of constructs. Each kit contains everything you need to perform 10 standard protein expression reactions of 140 µl volume in microtubes.

Proteins expressed


Standard soluble proteinsFab antibodies proteinsDisulfide-bonded proteins6–24 h / 30 µgBatchRTS Linear Template Kit PlusRTS Linear Template Fab KitRTS pIX3.0 VectorRTS pIVEX E. coli His-tag, 2nd Gen. V.

BR1400601

BR1400602




RTS 100/500 E. coli Disulfide Kits

FEATURES• Optimized reaction conditions for the expression of functional disulfide-bonded proteins• No need for refolding aggregated proteins• CECF technology for yields ables of up to 2.5 mg

APPLICATIONS• Expression of active proteins containing multiple disulfide bonds

DESCRIPTIONThe biotechrabbit RTS 100/500 E. coli Disulfide Kits provide an optimized combination of lysate, reagents, cha-perones, and buffer systems to produce maximum yields of active proteins containing multiple disulfide-bonds in a CECF system.The kit employs several innovative technologies:• Optimized high-yield (HY) E. coli lysate• Redox buffer to maintain the system under oxidizing conditions, which enhances disulfide-bond formation• Disulfide isomerase for rearrangement of disulfide bonds, which enhances the yields of correctly folded proteins

The CECF technology prolongs the reaction time up to 24 hours and, therefore, enhances protein yield. Transcripti-on, translation and disulfide-bond formation take place in the reaction compartment of the device. Substrates and energy components needed for a sustained reaction are continuously supplied via a semipermeable membrane. At the same time, potentially inhibitory interactions with byproducts are diluted by diffusion through the same mem-brane into the 1 ml feeding chamber. Depending on the particular protein, expression of up to several milligrams of protein per reaction can be obtained (e.g., >2 mg/ml of active urokinase).

Proteins expressed


Standard soluble proteinsDisulfide-bonded proteins(Fab antibodies)6–24 h / 80 µg (50 µl), 2.5 mg (1 ml)CECFRTS pIX3.0 VectorRTS pIVEX E. coli His-tag, 2nd Gen. V.

RTS 100 E. coli Disulfide KitBR1400401 24 rxn

of 50 µlReagents for coupled transcription/translation reactions


RTS 500 E. coli Disulfide KitBR1400501 5 rxn

of 1 mlReagents for coupled transcription/translation reactions

65


RTS 100 Wheat Germ Kit

FEATURES• Efficient expression of up to 5 µg protein per reaction• Compatible with non-optimized, pro- and eukaryotic T7 promoter-based vectors as well as linear templates• Excellent homogeneity of large proteins (264 kDa expressed)• No endotoxins, allows direct use in cell-based assays• Upscaling using high yield CECF wheat germ kits

APPLICATIONS• Fast parallel synthesis for screening purposes• Synthesis of mammalian or other proteins for functional testing without the disturbing endogenous protein background that is typical for in vivo expression• Synthesis of proteins dependent on eukaryotic chaperones

DESCRIPTIONThe biotechrabbit RTS 100 Wheat Germ Kit is a batch screening system allowing rapid protein expression within 2.5 hours. The kit allows use of a broad range of non-op-timized T7 promoter-based expression vectors from prokaryotic and eukaryotic origin in addition to established cell-free wheat germ vectors or linear templates. Using this approach, multiple constructs — including vectors that have not been optimized for wheat germ — can be scree-ned rapidly for expression and functionality in parallel. The procedure saves precious time otherwise required for template generation. The new batch system yields up to 100 µg protein per ml reaction, which is sufficient for many downstream applications.

Proteins expressed


Standard soluble proteinsComplex and large proteinsMembrane proteins3 h / 5 µgBatchRTS Wheat Germ LinTempGen SetRTS pIVEX Wheat Germ His6-tag V.

BR1402501 24 rxn of 50 µl



RTS 100/500 Wheat Germ CECF Kits

FEATURES• Successful expression of eukaryotic proteins , generally without sequence optimization• Synthesis of up to 1 mg functional eukaryotic protein• Maximized production of active heat-sensitive proteins, due to an optimum reaction temperature of 24°C

Proteins expressed


Standard soluble proteinsComplex and large proteinsMembrane proteins6–24 h / 50 µg (50 µl), 1 mg (1 ml)CECFRTS Wheat Germ LinTempGen SetRTS pIVEX Wheat Germ His6-tag V.

RTS 100 Wheat Germ CECF KitBR1401001 24 rxn

of 50 µlReagents for coupled transcription/translation reactions


RTS 500 Wheat Germ CECF KitBR1401101 5 rxn

of 1 mlReagents for coupled transcription/translation reactions

APPLICATIONS• Up to milligram-scale protein expression of functional eukaryotic proteins• Expression of multiple proteins in parallel

DESCRIPTIONThe biotechrabbit RTS 100/500 Wheat Germ CECF Kits enable high success rates for expression and solubili-ty, particularly for eukaryotic target proteins. The use of PCR-generated linear templates greatly improves convenience. The kits allow synthesis of proteins in a wide molecular weight range (up to 220 kDa, as tested), as well as synthesis of truncated protein variants (e.g., for epitope or functional domain mapping). While the RTS 100 Wheat Germ CECF Kit is designed for high-throughput protein expression, the RTS 500 version ensures higher yields.

The system can be used with linear templates as well as mRNA, vectors and plasmid DNA. Depending on the protein, expression yields of more than 1 mg protein per reaction can be obtained within 24 hours in a 1 ml reaction.


RTS 100 Insect Disulfide Kit

FEATURES• Eukaryotic Spodoptera frugiperda insect cell system• Endotoxin-free protein expression into microsomes• Generation of intra- and intermolecular disulfide bonds• Compatibility with linear templates and plasmids• Translocation into endoplasmic-reticulum-derived microsomes with optional glycosylation

APPLICATIONS• Expression and rapid testing of of antibody fragments (Fab, scFv) and disulfide proteins

DESCRIPTIONThe biotechrabbit RTS 100 Insect Disulfide Kit is designed for functional screening of antibody fragments and disulfi-de proteins. The reactions are free of endotoxins and can be applied directly in cellular assays.The kit uses a lysate containing endoplasmic-reticu-lum-derived microsomes harboring chaperones and enzymatic disulfide isomerase activity for the synthesis of proteins with intra- and intermolecular disulfide bonds and allowing optional glycosylation.The RTS 100 Insect Disulfide Kit is compatible with both linear templates and expression plasmids.

Proteins expressed


Standard soluble proteinsComplex and large proteinsDisulfide-bonded proteinsFab antibodiesGlycosylated proteinsMembrane proteins4 h / 2 µgBatchRTS Linear Template Kit PlusRTS Linear Template Fab KitRTS pIX4.0 Insect VectorRTS pIX3.0 VectorRTS pIVEX Wheat Germ His6-tag V.RTS pIVEX E. coli His-tag, 2nd Gen. V. RTS 100 E. coli LinTempGenSet, His

BR1401401

BR1401402




RTS 100 Insect Membrane Kit

FEATURES• Robust expression of a wide range of proteins (clotting factors, protein kinases, transcription factors)• Yields of up to 50 µg/ml protein, sufficient for functional studies or screening procedures• Compatibility with linear templates and plasmids• Expression screening in small volumes (>5 µl)Proteins expressed


Standard soluble proteinsComplex and large proteinsGlycosylated proteinsMembrane proteins4 h / 2 µgBatchRTS Linear Template Kit PlusRTS Linear Template Fab KitRTS pIX4.0 Insect VectorRTS pIX3.0 VectorRTS pIVEX Wheat Germ His6-tag V.RTS pIVEX E. coli His-tag, 2nd Gen. V.RTS 100 E. coli LinTempGenSet, His

BR1401501

BR1401502


Reagents for linked transcription/translation reactionsReagents for linked transcription/translation reactions


APPLICATIONS• Expression of membrane proteins without the need for additives• Synthesis of post-translationally modified eukaryotic proteins

DESCRIPTIONThe biotechrabbit RTS 100 Insect Membrane Kit was the first commercially available in vitro protein synthesis kit based on cell lysates derived exclusively from insect cell cultures. These homogeneous lysates from a Spodop-tera frugiperda cell line enable efficient expression of a broad range of eukaryotic proteins with posttranslational modifications. Now, with an optimized lysate production procedure, this kit is even more efficient.In addition, the lysates contain functional organelle mem-brane fractions, whose activity is required for posttrans-lational modification of eukaryotic proteins, including membrane proteins.RTS 100 Insect Membrane Kit can be used with linear templates as well as expression vectors.


RTS Amino Acid Sampler

FEATURES• Set of 20 individual amino acids, allowing amino-acid specific protein labeling for structure determination

APPLICATIONS• For use with RTS 100, RTS 500 and RTS 9000 E. coli kits• Ideal for labeling of proteins for NMR and X-ray analyses

DESCRIPTIONUsing RTS 500 or RTS 9000 E. coli HY Kits, protein yields are sufficient in many cases for subsequent use in NMR spectroscopy. The RTS Amino Acid Sampler provides ready-to-use stock solutions of amino acids which can be labeled and used to replace the amino acids provided in the RTS E. coli HY Kits.

BR1401801 5 rxn of 1 ml

20 L-amino acids (1 vial each)5 × DTT (1,3-Dithiothreitol)


RTS Solubility Enhancer Set

FEATURES• Compatible with RTS 100 E. coli HY Kit and RTS 100 Wheat Germ Kit• Includes five enhancers for up to 1.2 ml synthesis reaction, each

APPLICATIONS• Optimization of soluble yield of cell-free synthesized protein• Solubility enhancement of cytosolic, segregated and membrane proteins• Embedding of membrane proteins in detergent micelles• Solubilization of cofactors to be added to protein synthesis reactions

DESCRIPTIONThe biotechrabbit RTS Solubility Enhancer contains detergent, compatible solute and buffer components to increase the synthesis rate of soluble proteins. Membrane proteins can be embedded in detergent micelles during or after translation. Compatible solute and buffer are provided for improved folding of cytosolic and segregated proteins. The RTS Solubility Enhancer complements pro-tein folding measures for targets that cannot be sufficiently folded using chaperones. Each enhancer can be applied to up to 1.2 ml synthesis reactions using RTS 100 E. coli HY Kit and RTS 100 Wheat Germ Kit.

BR1402801 5 Enhancer D-Sorbitol, Solvent buffer, Brij35, DDM, DHPC



RTS DnaK Supplement

FEATURES• E. coli chaperones to enhance correct folding and increase solubility, even of large proteins• Compatible with RTS 100, 500, 9000 E. coli kits

APPLICATIONS• Enhance the expression of soluble proteins• Improved synthesis of functionally activity proteins

DESCRIPTIONThe Rapid Translation System (RTS) is an “open” cell-free protein expression system, in which reaction conditions can easily be adapted in a protein-specific manner by adding co-factors, ligands, detergents, or chaperones to the reaction mixture.The biotechrabbit RTS DnaK Supplement includes DnaK (heat shock protein 70, hsp 70), one of the key chaperone systems in E. coli, in combination with DnaJ (hsp 40) and GrpE . The RTS DnaK Supplement can be used to increa-se the amount of correctly folded and/or soluble proteins expressed in the RTS.DnaK can be used with proteins of any size and serves as an aggregation inhibitor that binds unfolded protein or transfers the protein to other chaperones, such as the GroE or the Hsp90 systems. The RTS DnaK Supplement can be used with the RTS E. coli HY platform.

BR1401601 5 rxn of 1 ml

Mix of DnaK, DnaJ and GrpEEnergy Mix (contains ATP)


RTS GroE Supplement

FEATURES• E. coli chaperone mix to enhance correct folding and increase solubility of expressed proteins• Compatible with RTS 100, 500, 9000 E. coli platforms

APPLICATIONS• Improved synthesis of functionally active proteins

DESCRIPTIONThe Rapid Translation System (RTS) is an “open” cell-free protein expression system, in which reaction conditions can easily be adapted in a protein-specific manner by adding co-factors, ligands, detergents, or chaperones to the reaction mixture.The biotechrabbit RTS GroE Supplement provides the GroEL and GroES (together referred to as GroE), which are cytoplasmic chaperones of E. coli reported to be involved in the de novo folding of 10–30 % of all cytoplas-mic proteins. It is not possible to predict that a particular protein is a substrate for GroE, GroEL or GroES. However, the presence of GroE during cellular or cell-free protein synthesis has been reported to lead to higher amounts of correctly folded and active product in many cases. The RTS GroE Supplement can be used with the RTS E. coli HY platform.

BR1401701 5 rxn of 1 ml

GroE Supplement (GroEL, GroES)Package contentCat. no. Size


Protein Synthesis for Site-Directed Labeling

RTS 100 Site-Specific Label Kit

Proteins with unique functional groups such as affinity labels, fluorescence moieties or drug conjugates offer high potential in many biotechnological or biomedical applications.An attractive alternative to known conventional posttrans-lational methods of protein modification is the site-directed incorporation of unnatural amino acids. In a specialized cell-free system, amino acids labeled with dyes or chemoselective reactive amino acids are incorporated stoichiometrically at defined positions of the expressed protein. Such homogenous site specific-modified proteins are the basis for a broad range of downstream applications.

• Protein-drug conjugates

• Stoichiometric incorporation of dyes, PEG

or other functionality

• Labeling with a broad range of commercially

available chemoselective reagents

FEATURES• Efficient incorporation of chemoselective reactive amino acids• High-yield synthesis of proteins with unnatural amino acids • No protein purification necessary prior to labeling• Minimal change of native protein structure• Stoichiometric incorporation (1 label per 1 protein)• Amber suppression-based expansion of the genetic code

Proteins expressed


Standard soluble proteinsDisulfide-bonded proteinsMembrane proteins1–24 h / 150 µgCECFRTS Linear Template Kit PlusRTS Linear Template Fab KitRTS 100 E. coli LinTempGenSet, HisRTS pIX3.0 VectorRTS pIVEX E. coli His-tag, 2nd Gen. V.

BR1402601 24 rxn of 50 µl


Package contentCat. no. Size APPLICATIONS• Synthesis of proteins containing a site specifically incorporated chemoselective reactive amino acid• Site-directed incorporation of dyes, PEG or other labels• Labeling with commercially available reagents• Labeled proteins for fluorescence-based applications such as microscale thermophoresis

DESCRIPTIONThe biotechrabbit RTS 100 Site-Specific Label Kit contains a modified E. coli lysate to drive coupled in vitro transcription and translation reactions especially for in-corporation of unnatural amino acids. The kit is compatible with both linear templates and plasmids, enabling fast and easy screening of protein modifications and expression optimization.Due to the optimized lysate and protocol, the RTS 100 Site-Specific Label Kit generates proteins with efficient incorporation of chemoselective reactive amino acid even at the C-terminus. The system uses an amber suppres-sion-based expansion of the genetic code. Minimal change of the native protein structure and stoichiometric incorporation of reactivity provide an enhanced tool for all downstream applications using conjugated proteins. Final labeling can be performed without prior purification of the protein; only buffer exchange is needed.

Cell-free protein synthesis Site-directed protein labeling

CR-AAChemoselective reactive amino acid

(CR-AA)

Chemoselectivereaction

Activated protein

Cell-freeexpression

UAA (stop)

CR-AA

Template Labeled protein

Ambersuppressor

tRNA


Linear Templates and Vectors for Cell-Free Protein Synthesis

biotechrabbit cell-free expression kits can be used with linear templates and expression vectors to produce the proteins of interest. The ability to use linear templates is one of the major advantages of cell-free compared to cell-ba-sed expression systems. Linear expression templates can be rapidly generated by PCR, allowing multiple constructs to be screened quickly and easily.Expression vectors enable protein production to be up-scaled. Either the genes of interest are directly inserted into an expression vector or linear templates are generated first by PCR and subsequently cloned. Protein yields achie-ved are sufficient for extended functional and structural analysis, or production of labeled proteins for NMR spec-troscopic or X-ray crystallographic studies.

• Fast PCR-based generation of linear templates

• Rapid expression screening and optimization

• Optimized expression vectors for scaling

production up to 6 mg per ml

• Flexible positioning of purification tags

1

2

3

4

5

6

7

8

9

10

No. Product Protein tag Expression kits ( see page 61 )

RTS Linear Template Kit Plus

RTS Wheat Germ LinTempGen Set, His6-tag

EasyXpress Linear Template Kit Plus

RTS Linear Template Fab Kit

RTS 100 E. coli LinTempGen Set, His-tag

RTS pIX4.0 Insect Vector

RTS pIX3.0 Vector

RTS pIVEX Wheat Germ His6-tag Vector Set

RTS pIVEX E. coli His-tag, 2nd Gen. Vector Set

EasyXpress pIX3.0 Vector

PCR generated linear templates

His-tag, Strep-tag, Signal peptide

His-tag

His-tag, Strep-tag, Signal peptide

His-tag, Signalpeptide

His-tag

Tag from cloned gene

Tag from linear template

His-tag

His-tag

Tag from linear template

RTS 100 E. coli HY and Fab KitsRTS 100 Insect Disulfide and Membrane KitsRTS 100 Site-Specific Label Kit

RTS 100/500 Wheat Germ Kits

EasyXpress E. coli KitsRTS 100 Insect Disulfide and Membrane Kits

RTS 100 E. coli HY and Fab KitsRTS 100 Insect Disulfide and Membrane KitsRTS 100 Site-Specific Label Kit

RTS 100 E. coli HY KitRTS 100 Insect Disulfide and Membrane KitsRTS 100 Site-Specific Label Kit

RTS 100 Insect Disulfide and Membrane Kits

RTS 100/500/9000 E. coli HY and Fab KitsRTS 100/500 E. coli Disulfide KitsRTS 100 Insect Disulfide and Membrane KitsRTS 100 Site-Specific Label Kit

RTS 100/500 Wheat Germ Kits

RTS 100/500/9000 E. coli HY and Fab KitsRTS 100/500 E. coli Disulfide KitsRTS 100 Insect Disulfide and Membrane KitsRTS 100 Site-Specific Label Kit

EasyXpress E. coli KitsRTS 100 Insect Disulfide and Membrane Kits

T7 vectors for cloning genes or linear templates


RTS Linear Template Kit Plus

FEATURES• Efficient in vitro expression (E. coli and insect kits)• Primers for affinity tags (His6-tag /Strep-tag II)• Signal peptide sequence primers for expression translocation into microsomes (insect systems)

APPLICATIONS• Fast generation of multiple template constructs• Accelerated screening of protein expression templates and conditions

DESCRIPTIONThe RTS Linear Template Kit Plus accelerates the gene-ration of multiple templates for parallel protein expression. Constructs are suitable for expression in E. coli and insect cell-free lysates. Due to compatible restriction sites in the adapter primers, products can be easily cloned into pIX3.0 vectors for large-scale in vitro expression.The kit contains primers providing regulatory elements re-quired for optimal transcription and translation in cell-free expression systems. Specially designed 5'-untranslated regions reduce the formation of secondary structure in the translation initiation region, one of the most common causes of low expression rates. A His-tag or Strep-tag II can be added to either terminus, greatly simplifying protein purification and detection.A signal sequence enables expression of functional proteins containing disulfide-bonds or membrane proteins in RTS 100 Insect Membrane or Disulfide kits. A signal peptide can be introduced using the N-terminal primer, which contains a melittin sequence. This enables protein translocation into microsomal vesicles derived from insect cell endoplasmic reticulum.

Protein tagExpression kits

His-tag, Strep-tag, Signal peptideRTS 100 E. coli HY KitRTS 100 E. coli Fab KitRTS 100 Insect Disulfide KitRTS 100 Insect Membrane KitRTS 100 Site-Specific Label Kit

BR1402401 20 PCRs of 25 µl

PCR reagents for linear template generation


RTS Wheat Germ LinTempGen Set, His6-tag

FEATURES• Optimized spacing of eukaryotic regulatory elements• Time saving, no laborious cloning• Flexible positioning of His6-tag at the C- or N-terminus• Primary PCR product can be cloned into pIVEX vectors

APPLICATIONS• Rapid generation of linear expression constructs for screening applications• Linear template generation for optimized His6-tagged protein expression

DESCRIPTIONThe RTS Wheat Germ LinTempGen Set, His6-tag can be used to rapidly generate linear expression constructs that are ready for use with RTS Wheat Germ kits. PCR gene-rated linear templates allow fast protein expression from wild type or modified DNA sequences. Use PCR-based template generation to:• Add epitope-tag sequences for antibody detection• Introduce mutations• Change codon usage• Generate truncated proteins

Using linear templates for protein expression avoids the time-consuming step of cloning into expression plasmids and can be used for initial, rapid screening for expression.Gene-specific primers can be used for RTS Wheat Germ and E. coli LinTempGen sets enabling easy transfer of pro-ducts from any expression system to both the RTS Wheat Germ and E. coli platforms.For expression scale-up, the linear templates can be cloned into the RTS pIVEX Wheat Germ vector and used with RTS 100/500 Wheat Germ CECF kits.


His-tagRTS 100 Wheat Germ KitRTS 100 Wheat Germ CECF KitRTS 500 Wheat Germ CECF Kit

BR1401201 96 PCRs of 50 µl




EasyXpress Linear Template Kit Plus

FEATURES• Efficient in vitro expression in E. coli and insect kits• Primers encoding affinity tags (Strep-tag II/His6-tag)• Signal-peptide primers for translocation into microsomes (incest kits)

APPLICATIONS• Fast generation of multiple template constructs• Accelerated screening of protein expression templates and conditions

DESCRIPTIONEasyXpress Linear Template Kit Plus accelerates the ge-neration of multiple templates for parallel protein expres-sion. Constructs are suitable for expression in E. coli and insect cell-free lysates. Due to compatible restriction sites in the adapter primers, products can be easily cloned into pIX3.0 vectors for large-scale in vitro expression.The kit contains primers providing regulatory elements re-quired for optimal transcription and translation in cell-free expression systems. Specially designed 5'-untranslated regions reduce the formation of secondary structure in the translation initiation region, one of the most common causes of low expression rates. A His-tag or Strep-tag II can be added to either terminus, greatly simplifying protein purification and detection.A signal sequence enables expression of functional proteins containing disulfide-bonds or membrane proteins in RTS 100 Insect Membrane or Disulfide kits. A signal peptide can be introduced using the N-terminal primer, which contains a melittin sequence. This enables protein translocation into microsomal vesicles derived from insect cell endoplasmic reticulum.


His-tag, Strep-tag, Signal peptideEasyXpress E. coli KitsRTS 100 Insect Disulfide KitRTS 100 Insect Membrane Kit

BR1402101 20 PCRs of 25 µl



RTS Linear Template Fab Kit

FEATURES• Introduction of an affinity tag for protein purification• Signal peptide sequence for translocation into microsomes• Compatibility with E. coli HY, Fab and Insect kits

APPLICATIONS• PCR-based inear templates for protein expression in E. coli and insect disulfide kits• Fast template generation for expression of Fab and scFv antibody fragments

DESCRIPTIONThe RTS Linear Template Fab Kit is designed for rapid PCR-based generation of templates directly used in the E. coli Fab and Insect kits allowing functional screening of antibody fragments and disulfide proteins and to facilita-te the assignment of biological functions to genes. The templates can be generated with optional affinity tag and/or signal peptide sequences for protein purification and translocation into microsomes.

The RTS Linear Template Fab Kit generates templates with elements to drive efficient coupled in vitro transcrip-tion and translation reactions for the synthesis of proteins with intra- and intermolecular disulfide bonds. The kit is the product of choice for functional screening of disulfide proteins and optimization of constructs. One kit contains sufficient volumes of primer solutions to perform 96 stan-dard protein expression reactions with reaction volumes of 25 µl.


His-tag, Signal-peptideRTS 100 E. coli HY KitRTS 100 E. coli Fab Kit RTS 100 Insect Disulfide KitRTS 100 Insect Membrane KitRTS 100 Site-Specific Label Kit

BR1402201 96 PCRs of 25 µl




RTS 100 E. coli LinTempGen Set, His-tag

DESCRIPTIONThe RTS 100 E. coli LinTempGen Set is designed to rapidly generate linear expression constructs that are ready for use with RTS 100 E. coli HY, Fab and insect kits.PCR generated linear templates allow fast protein ex-pression from wild type or modified DNA sequences. Use PCR-based template generation to:• Add epitope-tag sequences for antibody detection• Introduce mutations• Change codon usage• Generate truncated proteins

Using linear DNA templates for protein expression avoids the time-consuming step of cloning into expression plas-mids and can be used for initial, rapid screening for expres-sion.Gene-specific primers can be used for RTS Wheat Germ and E. coli LinTempGen sets enabling easy transfer of pro-ducts from any expression system to both the RTS Wheat Germ and E. coli platforms.For expression scale-up, the linear templates can be cloned into RTS pIVEX E. coli vectors and used with RTS 100/500/9000 E. coli CECF kits.

FEATURES• Contains all regulatory elements in optimal spacing• Option for templates with N- or C-terminal His6-tag• Primary PCR product can be cloned into pIVEX vectors• For use with RTS 100 E. coli HY and insect kits

APPLICATIONS• Fast gene template generation without cloning• Generate linear templates for rapid screening and optimization of His6-tagged protein expression


His-tagRTS 100 E. coli HY KitRTS 100 Insect Disulfide KitRTS 100 Insect Membrane KitRTS 100 Site-Specific Label Kit

BR1400801 96 PCRs of 50 µl



RTS pIX4.0 Insect Vector

BR1400901 1 vector 25 µg RTS pIX4.0 vector DNAPackage contentCat. no. Size


Tag from cloned geneRTS 100 Insect Disulfide KitRTS 100 Insect Membrane Kit

FEATURES• Regulatory elements for efficient transcription and translation• Multiple cloning site and translational stop codons in all reading frames• Optimized untranslated regions stabilizing and protecting RNA from degradation by exonucleases• RNA of defined length via optional plasmid linearization

APPLICATIONS• Cloning genes for efficient synthesis of proteins using the RTS 100 Insect kits

DESCRIPTIONThe RTS pIX4.0 Insect Vector is designed for cloning genes into its multiple cloning site. The vector contains effi-cient untranslated regions specific for the insect expres-sion system, a T7 transcription promoter and additional elements allowing high-yield expression using the RTS 100 Insect Membrane and kits.

The RTS pIX4.0 Insect Vector offers efficient initiation of translation through a T7 promoter element combined with an optimized 3' untranslated region, a T7 terminator, and an optimally positioned linearization site. The expression template is added to the in vitro transcription or coupled transcription/translation reactions.


RTS pIX3.0 Vector

FEATURES• Multiple cloning site compatible with linear templates generated with RTS Linear Template Kit Plus and RTS Linear Template Fab Kit• High copy origin of replication• Cell-free protein synthesis scale-up to after expression screening using linear templates• Free of any potentially interfering regulatory elements ensuring efficient transcription and translation• Compatible with RTS E. coli and insect kits

APPLICATIONS• Cloning of RTS Linear Templates for expression in RTS E. coli and insect kits

DESCRIPTIONThe cloning vector RTS pIX3.0 is designed for cloning linear templates generated with RTS Linear Template Plus and Fab kits to subsequently up-scale protein synthesis in E. coli batch systems to the CECF format or to use with RTS insect kits. The vector contains a multiple cloning site compatible with that of the linear templates.

The RTS pIX3.0 Vector enables high-level protein expres-sion using the RTS 100/500/9000 E. coli HY HY kits, E. coli Disulfide and RTS Insect kits. The kit exhibits the same performance as the RTS pIVEX His-tag, 2nd Gen. Vector Set, which is used for classic cloning.


Tag from linear templateRTS 100 E. coli HY KitRTS 500 ProteoMaster E. coli HY KitRTS 9000 E. coli HY KitRTS 100 E. coli Fab KitRTS 100 E. coli Disulfide KitRTS 500 E. coli Disulfide KitRTS 100 Insect Disulfide KitRTS 100 Insect Membrane KitRTS 100 Site-Specific Label Kit

BR1402701 1 vector 25 µg pIX3.0 vector DNAPackage contentCat. no. Size

RTS pIVEX Wheat Germ His6-tag Vector Set

FEATURES• Stable circular templates ensure high yields in preparative-scale expression reactions• For construction of C- and N-terminal His6-tag proteins

APPLICATIONS• Expression vector for preparative-scale expression in eukaryotic wheat germ lysates

DESCRIPTION

The RTS pIVEX Wheat Germ vectors are designed for high yield cell-free expression of His6-tagged eukaryotic proteins using the RTS Wheat Germ kits.

The two vectors contain all regulatory elements necessary for in vitro expression based on a combination of T7 RNA polymerase and wheat germ lysates. They enable the attachment of a His6-tag in different orientations, at the C- and N-terminus of the protein. Cloning into RTS pIVEX Wheat Germ His6-tag vectors allows optimal protein ex-pression in RTS Wheat Germ CECF kits to produce more than 1 mg protein per 1 ml synthesis volume in a 24-hour reaction.


His-tagRTS 100 Wheat Germ KitRTS 100 Wheat Germ CECF KitRTS 500 Wheat Germ CECF Kit

BR1401301 2 vectors 10 µg pIVEX 1.3 WG vector DNA10 µg pIVEX 1.4 WG vector DNA



RTS pIVEX E. coli His-tag, 2nd Gen. Vector Set

FEATURES• Stable circular templates to ensure high yields in preparative-scale expressions• For construction of C- and N-terminal His6-tagged proteins• Optimal spacing of all regulatory elements• For use with RTS E. coli and Insect kits

APPLICATIONS• Preparative-scale expression of His6-tagged proteins

DESCRIPTIONThe RTS pIVEX E. coli vectors are designed for high-yield cell-free expression of His6-tagged proteins using a broad range of RTS expression kits.

The two vectors contain all regulatory elements necessary for in vitro expression based on a combination of T7 RNA polymerase and RTS expression lysates. They enable the attachment of a His6-tag in different orientations at the C- and N-terminus of the protein. Cloning into RTS pIVEX His6-tag vectors allows optimal protein expression with high yields.


His-tagRTS 100 E. coli HY KitRTS 500 ProteoMaster E. coli HYRTS 9000 E. coli HY KitRTS 100 E. coli Fab KitRTS 100 E. coli Disulfide KitRTS 500 E. coli Disulfide KitRTS 100 Insect Disulfide KitRTS 100 Insect Membrane KitRTS 100 Site-Specific Label Kit

BR1400701 2 vectors 10 µg pIVEX2.3d vector DNA10 µg pIVEX2.4d vector DNA


EasyXpress pIX3.0 Vector

FEATURES• Multiple cloning site compatible with Linear Templates generated with EasyXpress Linear Template Plus, RTS Linear Template Plus and Fab kits• High copy origin of replication• Cell-free protein synthesis scale-up to after expression screening using linear templates• Free of any potentially interfering regulatory elements ensuring efficient transcription and translation• Compatible with RTS E. coli and insect kit series

APPLICATIONS• Cloning of RTS Linear Templates for expression in E. coli kits

DESCRIPTION

The cloning vector EasyXpress pIX3.0 is designed for clo-ning linear templates generated with EasyXpress Linear Template Plus, RTS Linear Template Plus and Fab kits to up-scale protein synthesis in E. coli-based systems. The vector is also compatible with the RTS Insect kit series. The vector contains a multiple cloning site compatible with that of the linear templates. The EasyXpress pIX3.0 Vector exhibits the same performance as the RTS pIVEX His-tag, 2nd Gen. Vector Set, which is used for classic cloning.


Tag from linear templateEasyXpress E. coli KitsRTS 100 Insect Disulfide KitRTS 100 Insect Membrane Kit

BR1402301 1 vector 25 µg pIX3.0 vector DNAPackage contentCat. no. Size


His-Tag Purification Kit

FEATURES• Optimized for purification of up to 10 mg His-tag protein from cell-free synthesis lysate • Convenient use, all components included in the kit• High-capacity His-tag protein binding

APPLICATIONS• Efficient purification of His-tagged proteins from 1 ml cell-free expression reactions

BR1002001

BR1002002

5 preps

25 preps

1.25 ml Ni-NTA Resin5 ml His-tag Binding Buffer20 ml His-tag Wash Buffer5 ml His-tag Elution Buffer5 Columns (empty)6 ml Ni-NTA Resin25 ml His-tag Binding Buffer100 ml His-tag Wash Buffer25 ml His-tag Elution Buffer25 Columns (empty)


DESCRIPTIONThe biotechrabbit His-Tag Purification Kit contains the Ni-NTA resin, empty columns and buffers needed for puri-fication of His-tag proteins. His-tags, the most commonly used recombinant protein tag, can be introduced with the cell-free linear template sets and expression vectors.The kit protocol is optimized to ensure best result when used with cell-free expression lysates. Just incubate the Ni-NTA resin with the cell-free expression reaction, transfer the mixture to the empty column, wash and elute the His-tag protein.

His-Tag Purification Mag-Bead Kit

FEATURES• Optimized for purifying up to 20 µg His-tag protein from small-scale cell-free synthesis lysates• Convenient use, all components included in the kit• Magnetic Ni-NTA beads for fast and easy handling• High-capacity His-tag protein binding

APPLICATIONS• Efficient purification of His-tagged proteins from 50 µl cell-free expression reactions

BR1002101

BR1002102

24 preps

96 preps

0.5 ml Ni-NTA Mag-Beads17 ml Ni-NTA Bead Binding Buffer15 ml Ni-NTA Bead Wash Buffer12 ml Ni-NTA Bead Elution Buffer2 ml Ni-NTA Mag-Beads68 ml Ni-NTA Bead Binding Buffer60 ml Ni-NTA Bead Wash Buffer48 ml Ni-NTA Bead Elution Buffer


DESCRIPTIONThe biotechrabbit His-Tag Purification Mag-Bead Kit is de-signed to purify His-tag proteins from small-scale cell-free expression reactions (50 µl). The kit contains Ni-NTA mag-netic beads and optimized buffers guaranteeing excellent recovery of His-tagged proteins from cell-free expression. The magnetic beads allow easy handling and are the tool of choice for purifications from small volumes.


High Affinity His-tag Protein Binding

Modern proteomics relies on reliable and sensitive detec-tion and labeling of tagged proteins, such as those bearing the 6xHis-tag. However, the low affinity of currently availa-ble chelators used for protein binding limits the sensitivity of many applications for selective, site-specific conjugation of proteins. To address this issue, biotechrabbit offers a superior His-tag protein ligand for the reversible binding of proteins and cell surfaces: Tris-NTA.

With a binding affinity that is approximately four orders of magnitude higher than monovalent metal ion chelators (such as nitrilotriacetic acid and iminodiacetic acid), Tris-NTA increases the sensitivity in western hybridization, biosensor analysis, functional protein analysis, protein purification and many other applications.

• 10,000 times higher binding affinity than

conventional chelators

• Reversible binding of biomolecules

• Great flexibility in functional assays

Tris-NTA Amine / Biotin

DESCRIPTIONHis-tags are one of the most commonly used tags for protein expression analysis. Conventional metal ion chelators, such as nitrilotriacetic acid (NTA) and iminodiacetic acid (IDA), bind His-tags with low affinities in the range of 10 µM. The biotechrabbit Tris-NTA complexes three NTA groups that together bind a 6×His-tag with an affinity that is four orders of magnitude higher (1 nM) than is possible with conventional chelators (10 µM). The binding of His-tags is stoichiometric and single-molecule detec-tion is possible. Binding is reversible: bound His-tags can be released with imidazole or ethylenediaminetetraacetic acid (EDTA).biotechrabbit Tris-NTA is available with a free amine group or conjugated to biotin. It can be used in a large range of applications, including protein detection and labeling, cou-pling proteins, lipids or cells to surfaces, protein purificati-on and reversible protein modification.Notice to purchaser: Limited License ( see page 79 )

Tris-NTA AmineBR1001101 100 µg 100 μl Tris-NTA Amine (1 mg/ml)


Tris-NTA BiotinBR1001201 100 µg 100 μl Tris-NTA Biotin (1 mg/ml)

FEATURES• A complex of three Ni-NTA groups ensures high-affinity binding of His-tags • Binding affinity is approximately four orders of magnitude higher than monovalent metal ion chelators• Protein binding is stoichiometric

APPLICATIONS• Reversible labeling of proteins or cell surfaces• Detection and analysis of target molecules• Immobilization of proteins, lipids and cells on surfaces• Purification and sample preparation of proteins • Coupling with microscopic or spectroscopic probes

Lata et al., J. Am. Chem. Soc., 2005, 127, 10205–10215

Tris-NTA Amine Tris-NTA Biotin

His-tagHis-tag

Tris-NTA 1 nM affinity mono-NTA 10 µM affinity

functionalunit

functionalunit

Protein Protein


NOTES


www.biotechrabbit.com© July 2015 biotechrabbit GmbH, all rights reserved. PTM-1005-001

Trademarksbiotechrabbit™, DirectUP™, GeneUP™, RevertUP™, UPzol™, StranDisplace™, UPstart™ (biotechrabbit GmbH); Agilent®, Bioanalyzer® (Agilent Technologies, Inc.); ROX™ (Applera Corporation or its subsidiaries); Beacon®, StepOne™, Viia® (Life Technologies); Bio-Rad®, CFX384™, CFX96™, Chromo4™, iCycler®, iQ™, MiniOpticon™, MyiQ™, Opticon™ (Bio-Rad Laboratories, Inc.); Cepheid®, SmartCycler® (Cepheid Corp.); Coomassie® (Imperial Chemical Industries); Eco™, Illumina® (Illumina Inc.); Eppendorf®, Mastercycler®, REALPLEX® (Eppendorf AG); LightCycler®, TaqMan® (Roche Group); Mx3000P®, Mx3005P®, Mx4000®, Stratagene® (Stratagene); QIAGEN®, Rotor-Gene® (QIAGEN GmbH); Quantica® (Jonathan Redfern); Scorpion® (DxS Limited); Takara®, Cycler Dice™ (Takara Shuzo Co. Ltd.); Techne® (Techne Corporation).

Legal Disclaimer and product use limitationbiotechrabbit GmbH makes every effort to include accurate and up-to-date information within this publication; however it is possible that omissions or errors might have occurred. Changes of publication can be made at any time without notice. All mentioned trademarks are protected by law. The purchase of biotechrabbit product does not include a license to perform any patented applications; therefore it is the sole responsibility of users to determine whether they may be required to engage a license agreement depending upon the particular application in which the products are used. biotechrabbit products are developed, manufactured, and sold for in-vitro use and laboratory use only. They are not suitable for administration to humans or animals. The products are to be used only by trained laboratory personnel who are familiar with potential hazards and in laboratories equipped to perform life science research. biotechrabbit products may contain chemicals which are harmful if misused. Due care should be taken with all products to avoid direct contact with components that are potentially harmful.

Tris-NTA Amine / BiotinThe use of this product for research purposes is covered by a license from QIAGEN Group. No rights to use this product to perform or offer diagnostic, companion diagnostic, commercial testing or other commercial services for money or money’s worth are granted by the supply of this product expressly, by implication or by estoppel. Should you wish to use this product for any other purpose not covered by this license, please contact QIAGEN Corporate Business Development at [email protected].

biotechrabbit GmbHNeuendorfstr. 24a16761 Hennigsdorf, GermanyPhone: +49 3302 20754 10Fax: +49 3302 20754 11

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Deutsche Bank BerlinBLZ: 10070024Konto Nr. 062889101BIC: DEUTDEDBBERIBAN: DE49100700240062889101

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