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SuperArray Bioscience Corporation June 2003 Updates

SuperArray Bioscience Corporation June 2003 Updates. New Product Lines: MultiGene-12™ RT-PCR Profiling Kits SureSilencing™ siRNA Kits. PCR & RT-PCR Related Products: MultiGene-12™ RT-PCR Expression Profiling Kits. New Array Verification and Gene Expression Profiling Methods from SuperArray.

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SuperArray Bioscience Corporation June 2003 Updates

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  1. SuperArray Bioscience Corporation June 2003 Updates New Product Lines: MultiGene-12™ RT-PCR Profiling Kits SureSilencing™ siRNA Kits

  2. PCR & RT-PCR Related Products:MultiGene-12™ RT-PCR Expression Profiling Kits New Array Verification and Gene Expression Profiling Methods from SuperArray

  3. Advantages of MultiGene-12™RT-PCR Profiling Kits • Pathway-specific sets of genes • Profiles eleven closely related genes in a single analysis • Processes up to eight experiments simultaneously • Ideal for multi-gene profiling and array verification • Contains carefully designed and tested primers • Provides a simple way to multiplex RT-PCR analyses

  4. MultiGene-12™ Kit Contents • Kit Contents: • PCR-1000A: ReactionReady™ PCR master mix 1000 • Lyophilized Powder for 96 Reactions (8 tubes, 12 reactions per tube) • Taq DNA Polymerase, Buffer, Nucleotides (dNTPs) • Gel loading dye for end point analysis by agarose gel electrophoresis • MultiGene-12™ Primer Strips: • Eight identical 12-well strips. Each tube represents one gene. • Eleven closely-related genes from one biological pathway • Twelfth tube contains GAPD for normalization

  5. Additional Materials Required Accessory Products (Sold Separately): C-01: ReactionReady™ First Strand cDNA Synthesis Kit* Enough reagents for 8 reverse transcription reactions Buffers RI, RE, P, BC, H2O * At this time, not available in all international territories. Or Individual Components from Promega: Reagent Product Random Primers Promega Catalog # C118 RNase Inhibitor Promega Catalog # N2511 Reverse Transcriptase Promega Catalog # M1701 dNTPs Promega Catalog # U1330

  6. Forward and Reverse Primer Sets for Individual Genes ReactionReady™ PCR master mix 1000 PCR-1000A Experimental RNA 1 2 3 4 5 6 7 8 aliquot Experiments PH-nnn or PM-nnn C-01 37 oC, 60 min ReactionReady™ First Strand cDNA Synthesis Kit* * At this time, not available in all international territories. PCR Overview of MultiGene-12™ RT-PCR Profiling Kit Procedure Gel Analysis

  7. Initial MultiGene-12™ Product Offering • MultiGene-12™ Human Caspase RT-PCR Profiling Kit PH-001A CASP 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 13 • MultiGene-12™ Human BCL2 RT-PCR Profiling Kit PH-002A BAD, BAK1, BAX, BCL2, BCL10, BCL2A1, BCL2L2, BCLxL, BCLxS, BNIP3, BIK • MultiGene-12™ Human Integrin-I RT-PCR Profiling Kit PH-003A ITGA 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 2B • MultiGene-12™ Human Integrin-II RT-PCR Profiling Kit PH-004A ITGA 11, ITGAL, ITGAM, ITGAV, ITGAX, ITGB1, 2, 3, 4, 5, 6

  8. Initial MultiGene-12™ Product Offering • MultiGene-12™ Human MMP-I RT-PCR Profiling Kit PH-005A MMP 1, 2, 3, 7, 8, 9, 10, 11, 12, 13, 14 • MultiGene-12™ Human MMP-II RT-PCR Profiling Kit PH-006A MMP 15, 16, 17, 20, 24, 26, MMPL1, TIMP1, 2, 3, 4 • MultiGene-12™ Human Cytokine-I RT-PCR Profiling Kit PH-007A IL1B, 2, 4, 5, 6, 8, 11, 12A, 13, 15 • MultiGene-12™ Human Cytokine-II RT-PCR Profiling Kit PH-008A IL16, 17, 18, 25, IFNG, IL1A, CSF2, MIF, TGFB1, 3, LTA

  9. MultiGene-12™ Products Coming Soon • Equivalent Products for Mouse • PM-001 through PM-008 • MultiGene-12™ Mouse RT-PCR Profiling Kit • Eight new products every month • Human and Mouse products released on alternate months

  10. TGFb1 TGFb3 GAPD CSF2 IL17 LTA IL1A IL16 IFNg MIF IL18 IL25 RNA from Jurkat cells: Stimulated With PMA and A23187 Unstimulated Differential Gene Expression Observed with a MultiGene-12™ RT-PCR Profiling Kit MultiGene-12™ Human Cytokine II RT-PCR Profiling Kit

  11. RNA Interference Products:SureSilencing™ siRNA Kits Knock down the expression of key genes in specific biological pathways

  12. The siRNA Concept Hutvagner and Zamore (2002) Curr. Opin. Genet. Dev.12: 225-232.

  13. T7 Promoter How we generate siRNA Populations PCR Gene-specific cDNA fragments Same ones used for our GEArrays T7 RNA Polymerase RNase III siRNA

  14. Negative Control siRNA SureSilencing™ siRNA LipofectAMINE2000™ (Invitrogen) transient transfection CELLS CELLS 24 to 48 h incubation Overview of SureSilencing™ siRNA Kit Procedure Harvest cells for RNA. Perform RT-PCR Verification. Verify decrease in message level of specific gene relative to negative control sample. Perform other biochemical, molecular or cell biological assay.

  15. Advantages of SureSilencing™ siRNA Kits • Knock down key genes in specific pathways • Pre-tested & validated. Over 70% suppression. • Ready to use for transfection • No enzymatic steps required • Includes negative control siRNA • Includes primers to verify down-regulated by RT-PCR

  16. SureSilencing™ siRNA Kit Contents • Kit Contents: • Gene-Specific siRNA population • To known down expression of gene by at least 70% • Enough for 90 transfections (24-well format) • Negative Control siRNA population • To control for non-specific effects • To normalize other assay results • Enough for 18 transfections (24-well format) • Gene-Specific Primer Pair • To verify decrease in gene expression • Enough for 100 polymerase chain reactions

  17. Additional Materials Required Accessory Products (Sold Separately): C-01: ReactionReady™ First Strand cDNA Synthesis Kit* Enough reagents for 8 reverse transcription reactions Buffers RI, RE, P, BC, H2O P-1000A: ReactionReady™ PCR master mix 1000 Enough lyophilized master mix for 96 reactions Eight tubes, twelve reactions each * At this time, not available in all international territories. Or Individual Components from Promega: Reagent Product Random Primers Promega Catalog # C118 RNase Inhibitor Promega Catalog # N2511 Reverse Transcriptase Promega Catalog # M1701 dNTPs Promega Catalog # U1330 Taq DNA Polymerase Promega Catalog # M2661

  18. Initial SureSilencing™ Product Offering • RH-0001 APC Adenomatosis polyposis coli • RH-0002 YY1 YY1 transcription factor • RH-0003 E2F1 E2F transcription factor 1 • RH-0004 BRCA1 Breast cancer 1, early onset • RH-0005 BAD BCL2-antagonist of cell death • RH-0006 MYC c-myc proto-oncogene • RH-0007 STAT1 Signal transducer and activator of transcription 1 • RH-0008 FADD Fas-associated via death domain • RH-0009 BCL2 B-cell CLL/lymphoma 2 • RH-0010 LCK Lymphocyte-Specific Protein Tyrosine Kinase • Product base could potential be extended to most of our gene list. • Priority assigned to key upstream regulators of biological pathways.

  19. SureSilencing™ Products Coming Soon • Product base could potential be extended to most of our gene list. • Priority assigned to key upstream regulators of biological pathways. • NFkB1 Nuclear factor of kappa light polypeptide gene enhancer in B-cells 1 • CDKN2A Cyclin-dependent kinase inhibitor 2A • EGFR Epidermal growth factor receptor • MAPK1 Mitogen-activated protein kinase 1

  20. GEArray™ cDNA Arrays Kits MultiGene-12™ RT-PCR Profiling Kits SureSilencing™ siRNA Kits Relationship Between SuperArray Product Lines

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