Judi Tilghman, Ph.D. Shelly Staat Product Manager Product ManagerGenomics and Proteomics Integrated Systems

2001 International Sales Meeting

2001 MultiPROBE II Product Focus

PackardBioScience

Increase visibility of MPII platformsin the genomics & proteomics market

Genomics

Developing co-marketing agreements with reagent vendorsValidated protocols on MPII, HT & Talon platforms

– Development work being done in-house, at vendor sites & customer sites

Application notes with joint customersProvide customer referencesProvide easy access to protocols, app notes and posters

from our website via reciprocal web links

PackardBioScience

Millipore

Montage Plasmid Miniprep

Montage PCR Cleanup

vacuum filtrationproprietary size exclusion filter plates

PackardBioScience

RNAture

Isolation of mRNA

96-Well Format Automation friendlyHigh-throughput capability

Fast Process

Easy Protocol3 steps standard protocolOne-step high-throughput

protocol

mRNA Express™ Kit

PackardBioScience

Prolynx

Utilizes proprietary super-paramagnetic particles (patent pending)

Particles bind dye terminators, not DNA extension products, “reverse purification”

No centrifuge rotors or carriers, or vacuum station

Particles are pre-loaded in a standard 96-well and 384-well formats

RapXtract DyeTerminator Removal Kit

PackardBioScience

Macherey-Nagel

silica membrane technology in combination with vacuum processing

fully automated, high-throughput purification of plasmid DNA & total RNA

high-throughput clean-up of PCR products

NucleoSpin® Robot Plasmid kit

NucleoSpin® Multi-96 ExtractNucleoSpin® Multi-96 RNA

NucleoSpin® Robot Plasmid kit

NucleoSpin® Multi-96 ExtractNucleoSpin® Multi-96 RNA

PackardBioScience

Genomics - Summary

Qiagen

– Validated protocols on MPII, HT, Talon platforms

• Qiaprep plasmid prep

– Development of PCR cleanup protocol in progress

– Application note in progress Millipore

– Validated protocols on MPII, HT platforms for

• Montage & MultiScreen plasmid prep & pcr cleanup

• Protocols on Talon platform in progress

• 2 application notes in progress

PackardBioScience

Genomics - Summary

RNAture

– mRNA express kit currently being developed in-house

Prolynx

– RapXtract kit currently being developed in-house

Macherey-Nagel

– Validated protocol for Nucleospin 96 plasmid prep kit

– Application note in progress

Future collaborations with:

– Promega, Sigma and Chemagen

PackardBioScience

Proteomics

The study of proteins expressed by genomes

Completion of the sequencing of the 1st draft of human genome indicates there are approximately 250,000 proteins in the human genome

Only 2-5% of proteins in human genome have been identified

Genome (DNA)Genome (DNA)

Transcriptome (mRNA)Transcriptome (mRNA)

Proteome (protein)Proteome (protein)

PackardBioScience

Increase visibility of MPII platformsin the genomics & proteomics market

ProteomicsDeveloping strategic marketing relationships with key players in

the proteomics market

– Micromass

– Applied Biosystems

– Bruker

– MilliporeOEM agreementsPromotion of MPII platforms as “stand alone” system for

Proteomics sample prep & spotting of MALDI targets

PackardBioScience

SampleSamplePrepPrep

2-D2-DGELS ORGELS OR

BLOTSBLOTS

MASS SPEC

MALDI-TOF

SpotSpotCutterCutter

ProteinProteinDigestDigestMALDI MALDI

SpottingSpotting

SampleSample

SWISS-PROT/ TrEMBL

ESIESIMS/MSMS/MS

ImagingImagingD/BASED/BASE

PepSeqPepSeq

The proteomic processSWISS PROT/

TrEMBL

MultiPROBEMultiPROBE

MicroMassABI

Bruker

MultiPROBEMultiPROBE

PackardBioScience

Digestion ProtocolDe-stain

100 mM ammonium bicarbonate and acetonitrile

DehydrateAcetonitrile

Reduce10 mM dithiothreitol in 100 mM ammonium bicarbonate, 30 minutes

Alkylate55 mM iodoacetamide in 100 mM ammonium bicarbonate, 20 minutes

Wash100 mM ammonium bicarbonate

DehydrateAcetonitrile

Digest6 ng/µL Trypsin in 50 mM ammonium bicarbonate (25 µL), 5 hours at 37ºC

Extract1% formic acid / 2% acetonitrile (30 µL)

Spot96-spot MALDI target (2 x 2.5 µL)

PackardBioScience

The proteomic process

ProteinProteinDigestDigest

MALDI MALDI SpottingSpotting

MASS SPECMALDI-TOF

MicromassABIBruker

MALDI Targets Range From 96 Grid on 2 X2 Format to 384 Grid on Standard Microplate FormatMultiPROBEMultiPROBE

PackardBioScience

Proteomics

PackardBioScience

Proteomics

PackardBioScience

Proteomics

Millipore ZipTips®

P10 size disposable tip which contains varying µg amounts of C18 Solid Phase

Tips are used to de-salt and concentrate extracted proteins prior to spotting on to a MALDI target plate

ZipTips with collars are currently available from Packard

Millipore will take over distribution within next 6 months

ZipTips will be packaged in “robotic friendly” Packard tip box

PackardBioScience

Proteomics - Summary

Micromass

– OEM with MPII standard

• Expand to include HT

• Forecast 70 - 80 units in 2001Applied Biosystems

– Strategic Marketing agreement in progress

• MPII, HT and Talon platformsMillipore

– ZipTip compatibility

– Developing automated platform for “vacuum based in-gel digest kit”

PackardBioScience

Digestion ProtocolDe-stain

100 mM ammonium bicarbonate and acetonitrile

DehydrateAcetonitrile

Reduce10 mM dithiothreitol in 100 mM ammonium bicarbonate, 30 minutes

Alkylate55 mM iodoacetamide in 100 mM ammonium bicarbonate, 20 minutes

Wash100 mM ammonium bicarbonate

DehydrateAcetonitrile

Digest6 ng/µL Trypsin in 50 mM ammonium bicarbonate (25 µL), 5 hours at 37ºC

Extract1% formic acid / 2% acetonitrile (30 µL)

Spot96-spot MALDI target (2 x 2.5 µL)

PackardBioScience

What does this mean for you?

More visibility in the proteomics market

= more MPII sales as stand alone systems