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EASY TO USE MICROARRAY PRODUCTS-TODAY!

EASY TO USE MICROARRAY PRODUCTS-TODAY!. Pat Mayer Global Product Manager MICROMAX Feb. 2002. PerkinElmer Microarray Solution. Spotters. Data Analysis Software. GeneTAC G3. Scanners. Glass Slides. PerkinElmer Genomics. Slide Scanning Service. NEN Brand Nucleotide

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EASY TO USE MICROARRAY PRODUCTS-TODAY!

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  1. EASY TO USE MICROARRAY PRODUCTS-TODAY! Pat Mayer Global Product Manager MICROMAX Feb. 2002

  2. PerkinElmer Microarray Solution Spotters Data Analysis Software GeneTAC G3 Scanners Glass Slides PerkinElmer Genomics Slide Scanning Service NEN Brand Nucleotide Analogs (Cyanine- and Biotin-labeled) GeneTAC Hybstation Labeling Kits Prespotted Microarrays Complete Microarray Systems

  3. PerkinElmer Microarray Solution

  4. Why MICROMAX Products? MICROMAX & Differential Gene Expression

  5. Why MICROMAX Products? Differential gene expression Gene expression Relative levels of RNA and proteins Differential gene expression Differences in levels of RNA and proteins as a result of changes in cellular phenotype

  6. Backgound MICROMAX Technology DNA Microarrays

  7. Backgound MICROMAX Technology Microarrays • A new and prominent technology that vies with PCR as the most significant commercial scientific advance of the decade. • Technology allows • Miniaturized format • Membrane or glass slide • Detection instrumentation • Analysis software

  8. Backgound MICROMAX Technology DNA Microarrays • Membrane Arrays • Oligonucleotide Arrays • cDNA or Long Oligo Arrays

  9. Backgound MICROMAX Technology Types of DNA Arrays • Membrane Arrays • cDNA spotted on nylon membrane • Rad nucleotides (chemiluminesence) • Gene expression and novel gene discovery • Advantages • Standard molecular biology techniques • Sensitive • Reusable • Unknown genes (ESTs) can be analyzed • Disadvantages • Single analysis per array • Low density • Radioactive safety

  10. Backgound MICROMAX Technology Types of DNA Arrays • Oligonucleotide chips • Oligo probes synthesized onto chip • Fluorescence detection • Gene expression and mutational analysis • Advantages • Very specific, can identify isotypic changes or SNPs • Tens of thousands oligos can be analyzed per chip • Automation • Disadvantages • Only known genes can be analyzed • Single analysis per array • Low sensitivity • Many spots make a gene • May be too specific for gene expression analysis • Not reusable

  11. Backgound MICROMAX Technology Types of DNA Arrays • Glass microarrays • cDNA or long oligos (50-70 mers) printed onto slides • Two-color fluorescence detection • Gene expression and novel gene discovery • Advantages • Dual detection for accurate differential analysis • Thousands of individual cDNA per slide • Unknown genes (ESTs) can be analyzed • Automation • Disadvantages • Low sensitivity • Not reusable

  12. Reverse Transcriptase *P* *P* *P* 33P dNTP cDNA mRNA Backgound MICROMAX Technology Labeling and detection for Microarrays Direct - radioactivity Preferred method for membrane arrays ** Single detection , Very sensitive**

  13. Reverse Transcriptase F F F fluoro- dNTP cDNA mRNA Backgound MICROMAX Technology Labeling and Detection for Microarrays Direct - fluorescence Standard method for glass and oligo arrays ** Dual detection, Good sensitivity**

  14. F F F F F F F F F F T T T T T T T T T T H H H Y Y Y Reverse Transcriptase B B B biotin-dNTP fluorescein-dNTP cDNA mRNA Backgound MICROMAX Technology Labeling and Detection forMICROMAX Indirect and amplified - Fluorescence Excellent method for glass arrays (MICROMAX-TSA) ** Dual detection, Excellent sensitivity**

  15. TSA Technology Review TSA

  16. TSA Technology Review • TSA is a proprietary, PATENTED technology developed at NEN • Patented under the name CARD (Catalyzed Reporter Deposition) • Commercialized under the name TSA(Tyramide Signal Amplification)

  17. TSA Technology Review Keys To TSA Inroduction of Horseradish Peroxidase (HRP) enzyme Labeled Tyramide

  18. TSA Technology Review SIGNAL AMPLIFICATION SIGNALAMPLIFICATION SIGNAL AMPLIFICATION

  19. TSA Technology Review SENSITIVITY SENSITIVITY SENSITIVITY

  20. TSA Technology Review TSA & Microarray Technology

  21. Signal Detection Amplify F F F F F F F F F F F F F H H H React with Tyramides Y Y Y T T T T T T T T T T T T T Incubate with HRP- conjugate Specific binding with hapten containing probe B B B Sample immobilized to a surface - slide • Used existing patented and proprietary technology TSA Tyramide Signal Amplification

  22. Simultaneous hybridization both Fluorescein and Biotin labeled total RNA Y Y Y Y Incubate with Anti-F-HRP Amplify with TSA-cyanine 3 Inactivate ResidualHRP Incubate with streptavidin-HRP Amplify with TSA-cyanine 5 TSA on Microarrays Sequential Fluorescence Deposition with TSA

  23. TSA Technology Review Reduced Customer Sample Requirements TSA reduces the input RNA sample up to 200-fold Increases opportunity for microarray technology use Tumor samples Reduced RNA sample costs Increased # of experiments

  24. MICROMAX microarray Analysis with Direct Fluorescence and Tyramide Signal Amplification 4800 Slide Direct Labeling Method TSA Labeling Method (100 µg Total RNA) (1 µ g Total RNA) TSA labeling method is very sensitive and needs as little as 0.5 to 1 µ gtotal RNA

  25. Direct and TSA Labeling: Scatter Plots Direct TSA Jurkat: Cyanine 3 Jurkat: Cyanine 3 HL-60: Cyanine 5 HL-60: Cyanine 5 TSA labeling method is highly sensitive and identifies more differentially regulated genes

  26. MICROMAX Products & Services

  27. MICROMAX Platforms Biotin/Fluorescein linked dNTPs Cyanine 3/5 dNTPs OR cDNA labeling isolate total RNA tissue samples imaging data analysis hybridize hybridize TSATM amplified fluorescence detection Fully integrated systems for streamlined application and consistent performance.

  28. MICROMAX Products and Services =Flexibility Microarray Products Reagent Kits Slide Scanning and Data Processing Services Cyanine 3- labeled dCTP, dUTP TSA Kit Direct Kit Cyanine 5- labeled dCTP, dUTP Complete Microarray Systems Quality Control microarray (Practice Slide: 25 genes) Human Screening microarrays Human Focused microarrays MICROMAXTM TSA Microarray Systems MICROMAXTM Direct Microarray Systems Glass Slides ULTRACLEAN I SUPERCHIP I Human Screening Array System I&II Human Array Systems I&II Human Focused Array Systems Chemical Labeling Kit

  29. MICROARRAY SLIDES • 2 human cDNA • microarrays • 2 practice arrays • coverslips • cDNA PREPARATION • Cyanine 3-dUTP • Cyanine 5-dUTP • Reverse Transcriptase • (RT)/RNase inhibitor • dNTP labeling mix • Unlabeled control RNA • HYBRIDIZATION • Hybridization Buffer INSTRUCTION MANUAL System Components MICROMAX Direct

  30. cDNA PREPARATION • Biotin and Fluorescein • Nucleotides • dNTP Nucleotide Mix • Reverse Transcriptase • (RT) • RNase Inhibitor • Unlabeled control RNA • MICROARRAY SLIDES • 2 human cDNA • microarrays • 2 practice arrays • coverslips • cDNA QC CHECK AND HYBRIDIZATION • Biotin- and Fluorescein-Control cDNAs • GeneScreenTM Membrane • Streptavidin-HRP and Anti-Fluor HRP • Hybridization Buffer • DUAL FLUORESCENCE • DETECTION • TSA Amplification Buffer • Cyanine 3-Tyramide • Cyanine 5-Tyramide • HRP Inactivation Reagent System Components MICROMAX TSA

  31. Platform Specifications MICROMAX Direct MICROMAX TSA • Sample Requirement • 20-80 ug Total RNA • Dynamic Range • 8-125,000 pg input mRNA (>104) • Differential Sensitivity •  2-fold ratio difference • Sample Requirement • 0.5-1 ug Total RNA • Dynamic Range • 0.04-200 pg input mRNA (>103.5) • Differential Sensitivity •  3-fold ratio difference

  32. MICROMAX Products & Services Complete MICROMAX Systems MICROMAX Human cDNA Microarray System I-Direct Direct Fluorescence Labeling System 5 slides MICROMAX Human cDNA Microarray System 1.1-TSA TSA Fluorescence Labeling System 5 slides MICROMAX Human cDNA Microarray System II-TSA TSA Fluorescence Labeling System 5 slides Positioning Everything for novice microarray user

  33. MICROMAX Products & Services MICROMAXReagent Kits MICROMAX Direct Reagent Kit Direct Fluorescence Labeling Kit 10 Labeling Reactions 50 Labeling Reactions MICROMAX TSA Labeling and Detection Kit TSA Fluorescence Labeling Kit 10 Labeling & Amplifcation Reactions 40 Labeling & Amplifcation Reactions Positioning Researchers who manufacture their own microarrays

  34. MICROMAX Products & Services MICROMAX Pre-SpottedMicroarrays MICROMAX Microarray: Human cDNA 1.1 (2400 human genes) MICROMAX Microarray: Human cDNA II (4800 human genes) MICROMAX Microarray: Quality Control Slides MICROMAX Microarray: Focused Array Slides (~270 human genes) 2 Microarrays GeneMAP Microarrays: Human, Mouse and Rat 5 microarrays Positioning Experienced researchers who don’t have access to cDNA arrays Experienced researchers who have their own labeling technology

  35. MICROMAX Products & Services MICROMAX Slide Scanning Service Scanning of all types of MICROMAX Slides Practice Arrays free of charge Slide Scanning Service Upgraded Web Based Service Positioning Novice researchers who don’t have access to fluorescent microarray scanner

  36. Selling GeneMAP Microarrays • What are they? • Spotted, preprinted arrays • Human, mouse and rat glass microarrays • Available Worldwide (non-exclusive) from PerkinElmer • Sales of GeneMAP will count towards your MM sales goals! • Under a different pac code however, 2063

  37. MICROMAX Competition COMPETITION

  38. MMICROMAX Competition Who sells “Complete Systems”? Labeling System Nucleotides Slides ONLY PERKINELMER!

  39. MICROMAX Competition • Competition Company Labeling Kits Slides Affymetrix x X Agilent X Clontech X X Amersham X Stratagene X X Genisphere X Mergen X HOMEBREW!!!! X X

  40. MICROMAX Competition Competing Technologies cDNA Labeling Technology AminoAllyl Clontech Dendrimer Genisphere HOMEBREW Slide Technology Short (20-30 mers) Oligo Affymetrix Mergen Medium (70-80 mers) Oligos Clontech cDNA Agilent Takara HOMEBREW

  41. MICROMAX Competition-Technologies Aminoallyl labeling technology……. Amino groups are incorporated during cDNA synthesis using aminoallyl-dUTP Cy3 or Cy5 monomer NHS ester is reacted with cDNA Labeling is independent of incorporation of dye-dNTP analogs ……is a novel technology which is cumbersome TARGET CUSTOMER FOR NEW CHEMICAL LABELING!

  42. MICROMAX Competition-Technologies Genisphere’ s dendrimer technology…... Labeling of cDNA RT with Unlabeled deoxynucleotides and Cy3™ or Cy5™ Primer Oligo (=3DNA Capture Sequence) Hybridize to Array Detect with 3DNA Fluor (Complementary Capture Sequence labeled with Cy-Dye™) …...only competing technology to MICROMAX-TSA Our customers have told us this doesn’t work very well-no specific signal

  43. Competitive Advantages • Low cost entry to microarray technology • Complete systems right out of the box • Broad product offering • License to do dual color fluorescence hybridization Every lab can afford it! Easy to use! Something for every lab! Two samples on one slide!

  44. MICROMAX Selling Probing Questions…... Are you currently using microarray technology in your research? Do you anticipate using microarray technology in the near future? Is your differential gene expression research limited by the amount of starting material? Are you just getting started in microarray technology? …….leads to providing customers with the right products

  45. QC Process for Microarray products • Labeling and Detection Kits • QC all components • Nucleotides HPLC QC • Nucleotides functional testing • Components used in functional testing of microarrays • Cross check in R & D and Applications Lab • Microarrays • Check PCR process • In process checks • Check autofluorescence of glass slides • Representative sequencing • Check after printing for spots • Functional testing of arrays • Spot morphology • Spot size • S/N • Cross check in R & D and Applications Lab

  46. New MICROMAX products in the last year • 4800 Arrays and Systems (1.1) (Alphagene genes) • Now manufacture our own arrays • TSA and Direct versions • Direct Kit • more economical-$40/slide • Focused Arrays • Human Apoptosis & Neurobiology • Additional Screening Arrays and System (II) • Incyte Genomics’ Gene Content • Coming Soon • Glass Slides for DNA spotting • New Chemical Direct Labeling System • Mouse and Rat Screening Arrays • Content from LION bioscience • Control Kits NEW

  47. Current MICROMAX Ads • MICROMAX TSA • “The NEN TSA Microarray Labeling and Detection Kit lets you do more with less” • Ran about 10 times in 2001-will be redesigned • MICROMAX Direct Kit Ad • “Simply the best microarray reagents on earth (or anywhere else you would care to explore!” • Ran about 2 times in 2001 and continue in 2002 • MICROMAX Microarray Ad • “A long time ago. . .in a galaxy far away. . .we started making pre-spotted microarrays. . .We Still Do.” • Newly created and will run in 2002 • MICROMAX Glass Slide Ad • “We have a new addition to the family. . .Introducing MICROMAX Glass Slides” • Newly created and will run in 2002 • MICROMAX Chemical Direct Labeling Ad • Not created yet and will run in 2002

  48. Introducing MICROMAX Glass Slides • ULTRACLEAN I • Glass Slides that are cleaned! • For homebrew coating procedures • Looking for evaluators (e-mail Pat) • MPS692 • SUPERCHIP I • Amino Coated Glass Slides • Looking for evaluators (e-mail Pat) • MPS695-5 slides • MPS696-20 slides Ultra-flat Low Intrinsic Fluorescence Bundling opportunities with PerkinElmer printers and spotters! Packaged in sets of 5 slides Ready to ship evals February 11, 2002 Ready to ship by end of February

  49. Breadth of Technical Support • In-field dedicated genomics specialists • Internal dedicated Technical Support • Leslie Stern X9117 • Internal dedicated Applications Lab • Testing and expanding microarray protocols • Research and Product Development Staff • Organized for efficiency and time to market improvements • External seminar series

  50. MICROMAX Sales Tools Available MICROMAX Seminar Will be updated ASAP to incorporate Packard instruments MICROMAX Literature Pieces Code Name of Piece H78255 MICROMAX Sell Sheet H78511 MICROMAX TSA BioTechniques Reprint P10208 MICROMAX Direct Reagent Kit Sell Sheet H78414 MICROMAX TSA Sell Sheet H78381 MICROMAX Focused Array Sell Sheet H78416 MICROMAX TSA Protocol Laminated Card Laminated card (H78416) is a must have for wet labs! Order at www.perkinelmerliterature.com

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