HIT Competent Cells Non-Heat Shock Transformation

Revolutionary 1-10 mins transformation to HIGH EFFICIENCY

HIT Competent Cells

Welcome to the world of HIT Competent CellsNon-Heat Shock Transformation

Only ICE

Prepare ice bucket, 37°C plating beads and selective plates. Thaw competent cell vial with room-temp. tap water or water bath for 10~20 seconds until 1/3 thawed.

Add DNA whose volume is less than 10% of volume of cells . Vortex 1 second.

Place on ice for 1-10 minutes

Plate Transfer onto 37°C dry selection plate media, spread using RBC plating beads.

Immediately incubate plate at 37°C (16-18 hours for HIT-DH5α, 8-24 hours for HIT-109).Observe growth of transformed colonies.

HIT Cells accept any circularised plasmids.

For reliable cloning into HIT Competent Cells

RBC TA Clonng Kit or RBC Repid Ligation Kit

HIT Non-Heat Shock Competent Cells are the fastest transforming cells worldwide.Due to our unique production process cells can be transformed to high efficiency in 1-10 minutes by simply placing on ice.HIT Competent Cells incorporate a wide range of strains and efficiencies for all your cloning and expression needs.Transform your work with HIT Non-Heat Shock Competent Cells.

Compared to Traditional Competent Cells

· No Heat Shock Required· Extremely Rapid Protocol· True 1 step/1 tube process· Guaranteed efficiency up to 109 cfu/ug plasmid· Economical for subcloning to library production· Increased Freeze Thaw resistance· Ideal for automation due to simplified protocol

RBC HIT Competent Cells should be stored in a -70˚C non frost-free freezer.Competent cells are sensitive to multiple cycles of freezing and thawing and exposure to temperature variations should be minimized.Each RBC HIT package comes with the HIT efficiency guarantee-HIT cells have been electronically temperature monitored to ensure -70˚C AT ALL TIMES during batch processing, shipping and storage.

HIT Competent Cells offer a powerful alternative to traditional competent cell preparations. Simply add DNA, vortex and place on ice for 1~10 min.Efficiency levels are equal or superior to traditional two hour multi-step protocols for commercial cells. Compared to average DIY competent cells, efficiency is 100-1000 times higher. No significant difference is observed for incubation within 1 to 10 min time frame.If cells are placed on ice > 30 min efficiency will be reduced 2~3 fold.

JUMBOValue 108High 108Super 109

EconomicalConvenient and EconomicalHigh Cloning EfficiencySuper Cloning Efficiency

Vector

T7 PromoterLacZα

APr ori

Multiple Cloning regionlacZα gene

r gene AP totototo

toto

T7 promoterM13 forward primerM13 reverse primer

4901491671

402359528

439375507

258511434

yyyyyyyII EEyyyyy IIICCEEEOOOOnnnnnllyyllyyyyOOOOOOOOOOOOOnn CCCCCCCCCCCEEEEEIIIOO llllllyyOOO CCAdditional Requirements : Only ICE HIT Competent Cells Protocol Clone With RBC TA Vector HIT Application Table (also see Genotype Table)

HIT Competent Cells show excellent stability in the presence of ligation buffer.Efficiency levels are basically equivalent within ligation buffer volume range of 1~10 μl.Traditional commercial competent cells are optimised for ligation buffer volumes of less than 5 μl.

HIT Competent Cells require warm plates and plating beads.Incubation at 37˚C prior to transformation is recommended in order to standardise conditions between platings. Room temperature plates and plating beads may be sufficient in certain cases.It is not recommended to use plates and plating beads directly from the refrigerator.

HIT Competent Cells require rapid thawing.This is very different from traditional chemical competent cells which always recommend a slow thawing process. The efficiency of fast thawing for HIT Competent Cells is 1~2 times that of slow thawing methods.

The above data indicates that HIT Competent Cells are very stable after freeze-thaw.In fact an initial freeze thaw may increase the efficiency slightly.It is not recommended to subject the cells to more than three freeze-thaw cycles.

Competent Cell Type HIT Competent Cells

Genotypes Applications

HIT-DH5α HIT-JM109 HIT-Blue HIT- 21

F- ( 80d lacZ M15) (lacZYA-argF)U169 hsdR17(r - m +) recA1 endA1 relA1 deoR

F'traD36proA+proB+ lacIq (lacZ)M15 l (lac-proAB) hsdR17 recA1 endA1 relA1

hsdR17(rk- mk+), recA1, endA1, gyrA96,thi-1,supE44,relA1,ac[F'proAB acIqZDM15Tn10(Tet )]

E.coli B, F-, dcm, ompT, hsdS(rB-mB-), gal (DE3)

end A Prevents plasmid degradation during extraction Yes Yes Yes No

recA Prevents DNA recombination Yes Yes Yes

hsd

Enhances transformation efficiency of selected PCR DNA strands and cDNA libraries

Yes Yes Yes Yes

deoREnhances transformation efficiency of high MW plasmids and cosmids

Yes No No No

LacZ M15 Blue-White screening Yes Yes Yes No

Lon Lon Protease Deficient, Improves Protein Yield

No No No Yes

ompT OmpT Protease Deficient, Improves Protein Yield No No No Yes

Speedy Thawing Handily

Effect of Plante Temperature

Less Freeze-Thaw Effect

Effect of Ligation Buffer Effect of Plating Temperature Comparison of Thawing Rate HIT Special FeaturesHITT SSppeecccciiiiiaaalll FFeaturesp ial Fe turesH TTSSSSSSppppppeeeeeecccciii F tComparison of Thawing RateC p awing Rarison of Tha ateCom HITTH TTTSSSS

HIT Competent Cells

RH717-J JUMBO Value 108 HIT-JM109 1 ml x 2 -70°C 8~10 hours growth, Blue / White Screening for Generation of cDNA libraries and subcloning -> 5X10 7 Transformants/ μg pUC19

RH717 JUMBO Value 108 HIT-JM109 100 μl x 10 -70°C 8~10 hours growth, Blue / White Screening for Generation of cDNA libraries and subcloning -> 5X10 7 Transformants/ μg pUC19

RH717-J80 JUMBO-80, Value 108 HIT-JM109 100 μl x 80 -70°C 8~10 hours growth, Blue / White Screening for Generation of cDNA libraries and subcloning -> 5X10 8 Transformants/ μg pUC19

RH718 High 108 HIT-JM109 100 μl x 10 -70°C 8~10 hours growth, Blue / White Screening for Generation of cDNA libraries and subcloning -> 1X10 8 Transformants/ μg pUC19

RH718-J80 JUMBO-80, High 108 HIT-JM109 100 μl x 80 -70°C 8~10 hours growth, Blue / White Screening for Generation of cDNA libraries and subcloning -> 1X10 8 Transformants/ μg pUC19

RH117-J JUMBO Value 108 HIT-Blue 1 ml x 2 -70°C General Cloning, Blue / White Screening, -> 5X10 7 Transformants/ μg pUC19

RH117 JUMBO Value 108 HIT-Blue 100 μl x 10 -70°C General Cloning, Blue / White Screening, -> 5X10 7 Transformants/ μg pUC19

RH117-J80 JUMBO-80, Value 108 HIT-Blue 100 μl x 80 -70°C General Cloning, Blue / White Screening, -> 5X10 7 Transformants/ μg pUC19

RH118 High 108 HIT-Blue 100 μl x 10 -70°C General Cloning, Blue / White Screening, libraries -> 1X10 8 Transformants/ μg pUC19

RH118-J80 JUMBO-80, High 108 HIT-Blue 100 μl x 80 -70°C General Cloning, Blue / White Screening, libraries -> 1X10 8 Transformants/ μg pUC19

RH119 Super109 HIT-Blue 100 μl x 10 -70°C General Cloning, Blue / White Screening, libraries -> 5X10 8 Transformants/ μg pUC19

RH217 Value 107 HIT-21 100 μl x 5 -70°C General Cloning protein expression -> 5X10 6 Transformants/ μg pUC19

RH217-J40 JUMBO-40 Value 107 HIT-21 100 μl x 40 -70°C General Cloning protein expression -> 5X10 6 Transformants/ μg pUC19

RG001 RBC Glass Plating Beads 100 g, 4mm RT

RH617-J JUMBO Value 108 HIT-DH5α 1 ml x 2 -70°C Blue / White Screening, General Cloning -> 5X10 7 Transformants/ μg pUC19

RH617 JUMBO Value 108 HIT-DH5α 100 μl x 10 -70°C Blue / White Screening, General Cloning -> 5X10 7 Transformants/ μg pUC19

RH617-J80 JUMBO-80, Value 108 HIT-DH5α 100 μl x 80 -70°C Blue / White Screening, General Cloning -> 5X10 7 Transformants/ μg pUC19

RH618 High 108 HIT-DH5α 100 μl x 10 -70°C Blue / White Screening for Generation of cDNA libraries and subcloning -> 1X10 8 Transformants/ μg pUC19

RH618-J80 JUMBO-80, High 108 HIT-DH5α 100 μl x 80 -70°C Blue / White Screening for Generation of cDNA libraries and subcloning -> 1X10 8 Transformants/ μg pUC19

RH619 Super108 HIT-DH5α 100 μl x 10 -70°C Blue / White Screening for Generation of cDNA libraries and subcloning -> 5X10 8 Transformants/ μg pUC19

CatNo. Product Specification Storeage Note

DH5α

Blue

JM10

921

Spread Competent Cells, 75~100 Plates

HIT Competent Cells Product Selection GuideThe above data indicates that HIT competent cells are very stable after freeze-thaw. In fact an initial freeze thaw may increase the efficiency slightly. It is not recommended to subject the cells to more than three freeze-thaw cycles.

The original transformation protocol dates back to 1970 (Mandel and Higa, J. Mol Biol 1970 , 53; 159). Just preparing the cells took two days. With a protocol two hours long, containing multiple steps, it was a revolution in molecular biology. Does this sound familiar? That's because it's still the most popular protocol in use today.

RBC HIT Competent Cells 1 Min competent cells are still currently the fastest transforming cells worldwide since it was launched in 2006. Due to our unique production process cells can be transformed to high efficiency in 1-10 minutes using a simple 1 step patented protocol. RBC provides a wide range of popular strains for all your cloning and expression needs. Transform your work with RBC HIT Competent Cells.

RBC HIT Competent Cell Efficiency GuaranteeRBC HIT Competent Cells should be stored in a -70°C non frost-free freezer. Competent cells are sensitive to cycles of freezing and thawing and exposure to temperature variations should be minimized. Each RBC HIT package comes with the HIT efficiency guarantee. HIT cells have been electronically temperature monitored to ensure -70°C AT ALL TIMES during batch processing, shipping and storage.

RBC HITCompetent Cell Advantage

Just Add Ice !Compared to traditional Commercial Competent Cells• Revolutionary 1 minute Transformation• -70°C Monitoring of competent cell shipments• True 1 step/1 tube process• Guaranteed efficiency up to 109 cfu/ug plasmid• Each batch efficiency checked during production• Economic pricing for regular and high efficiency cells• Ideal for bench users to automated cloning projects• Worldwide patent protectionHIT Competent Cell Selection DH5a, JM109, XL1-Blue, BL-21• JUMBO : Economical• VALUE 108 : Convenient and Economical• HIGH 108 : High Cloning Efficiency• SUPER 109 : Super Cloning Efficiency• HIT 96-Well : Cloning Libraries• JUMBO 80 : More Economical at Price/Prep

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