2001 Multi PROBE II Product Focus Judi Tilghman

2001 Multi. PROBE II Product Focus Judi Tilghman, Ph. D. Product Manager Genomics and Proteomics Shelly Staat Product Manager Integrated Systems 2001 International Sales Meeting

Increase visibility of MPII platforms in the genomics & proteomics market Genomics n Developing co-marketing agreements with reagent vendors u Validated protocols on MPII, HT & Talon platforms – Development work being done in-house, at vendor sites & customer sites u Application notes with joint customers u Provide customer references u Provide easy access to protocols, app notes and posters from our website via reciprocal web links Packard Bio. Science

Millipore Montage Plasmid Miniprep vacuum filtration proprietary size exclusion filter plates Montage PCR Cleanup Packard Bio. Science

RNAture m. RNA Express™ Kit n Isolation of m. RNA n 96 -Well Format u Automation friendly u High-throughput capability n Fast Process n Easy Protocol u 3 steps standard protocol u One-step highthroughput protocol Packard Bio. Science

Prolynx n Utilizes proprietary super- paramagnetic particles (patent pending) Rap. Xtract Dye Terminator Removal Kit n Particles bind dye terminators, not DNA extension products, “reverse purification” n No centrifuge rotors or carriers, or vacuum station n Particles are pre-loaded in a standard 96 -well and 384 -well formats Packard Bio. Science

Macherey-Nagel Nucleo. Spin® Robot Plasmid kit n silica membrane technology in combination with vacuum processing n fully automated, high -throughput purification of plasmid DNA & total RNA Nucleo. Spin® Multi-96 Extract Nucleo. Spin® Multi-96 RNA n high-throughput clean-up of PCR products Packard Bio. Science

Genomics - Summary u Qiagen – Validated protocols on MPII, HT, Talon platforms • Qiaprep plasmid prep – Development of PCR cleanup protocol in progress – Application note in progress u Millipore – Validated protocols on MPII, HT platforms for • Montage & Multi. Screen plasmid prep & pcr cleanup • Protocols on Talon platform in progress • 2 application notes in progress Packard Bio. Science

Genomics - Summary n RNAture – m. RNA express kit currently being developed inhouse n Prolynx – Rap. Xtract kit currently being developed in-house n Macherey-Nagel – Validated protocol for Nucleospin 96 plasmid prep kit – Application note in progress n Future collaborations with: – Promega, Sigma and Chemagen Packard Bio. Science

Proteomics Genome (DNA) n The study of proteins expressed by genomes n Completion of the sequencing of the 1 st draft of human genome indicates there approximately 250, 000 proteins in the human genome n Only 2 -5% of proteins in human genome have been identified Transcriptome (m. RNA) Proteome (protein) Packard Bio. Science

Increase visibility of MPII platforms in the genomics & proteomics market Proteomics u Developing strategic marketing relationships with key players in the proteomics market – Micromass – Applied Biosystems – Bruker – Millipore u OEM agreements u Promotion of MPII platforms as “stand alone” system for Proteomics sample prep & spotting of MALDI targets Packard Bio. Science

The proteomic process SWISS-PROT/ SWISS PROT/ Tr. EMBL D/BASE Imaging Pep. Seq Spot Cutter Sample Prep Multi. PROBE 2 -D GELS OR BLOTS Protein Digest MALDI Spotting Multi. PROBE MASS SPEC MALDI-TOF Micro. Mass ABI Bruker ESI MS/MS Packard Bio. Science

Digestion Protocol De-stain 100 m. M ammonium bicarbonate and acetonitrile Dehydrate Acetonitrile Reduce 10 m. M dithiothreitol in 100 m. M ammonium bicarbonate, 30 minutes Alkylate 55 m. M iodoacetamide in 100 m. M ammonium bicarbonate, 20 minutes Wash 100 m. M ammonium bicarbonate Dehydrate Acetonitrile Digest 6 ng/µL Trypsin in 50 m. M ammonium bicarbonate (25 µL), 5 hours at 37ºC Extract 1% formic acid / 2% acetonitrile (30 µL) Spot 96 -spot MALDI target (2 x 2. 5 µL) Packard Bio. Science

The proteomic process MASS SPEC MALDI-TOF Protein Digest MALDI Spotting Multi. PROBE Micromass ABI Bruker MALDI Targets Range From 96 Grid on 2 X 2 Format to 384 Grid on Standard Microplate Format Packard Bio. Science

Proteomics Packard Bio. Science

Proteomics Millipore Zip. Tips® n P 10 size disposable tip which contains varying µg amounts of C 18 Solid Phase n Tips are used to de-salt and concentrate extracted proteins prior to spotting on to a MALDI target plate n Zip. Tips with collars are currently available from Packard n Millipore will take over distribution within next 6 months n Zip. Tips will be packaged in “robotic friendly” Packard tip box Packard Bio. Science

Proteomics - Summary u Micromass – OEM with MPII standard • Expand to include HT • Forecast 70 - 80 units in 2001 u Applied Biosystems – Strategic Marketing agreement in progress • MPII, HT and Talon platforms u Millipore – Zip. Tip compatibility – Developing automated platform for “vacuum based in -gel digest kit” Packard Bio. Science

Digestion Protocol De-stain 100 m. M ammonium bicarbonate and acetonitrile Dehydrate Acetonitrile Reduce 10 m. M dithiothreitol in 100 m. M ammonium bicarbonate, 30 minutes Alkylate 55 m. M iodoacetamide in 100 m. M ammonium bicarbonate, 20 minutes Wash 100 m. M ammonium bicarbonate Dehydrate Acetonitrile Digest 6 ng/µL Trypsin in 50 m. M ammonium bicarbonate (25 µL), 5 hours at 37ºC Extract 1% formic acid / 2% acetonitrile (30 µL) Spot 96 -spot MALDI target (2 x 2. 5 µL) Packard Bio. Science

What does this mean for you? n More visibility in the proteomics market n = more MPII sales as stand alone systems Packard Bio. Science