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

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

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

Additional Materials RequiredAccessory Products (Sold Separately):

C-01: ReactionReady™ First Strand cDNA Synthesis Kit*Enough reagents for 8 reverse transcription reactionsBuffers RI, RE, P, BC, H2O

* At this time, not available in all international territories.

Or Individual Components from Promega:Reagent ProductRandom Primers Promega Catalog # C118RNase Inhibitor Promega Catalog # N2511Reverse Transcriptase Promega Catalog # M1701dNTPs Promega Catalog # U1330

Overview of MultiGene-12™ RT-PCR Profiling Kit Procedure

PCR

C-0137 oC, 60 min

ReactionReady™First Strand cDNA Synthesis Kit*

* At this time, not available in all international territories.

Experimental RNA12345678

aliquot

ReactionReady™ PCR master mix 1000PCR-1000A

Forward and Reverse Primer Sets for

Individual Genes

Exp

erim

ents

PH-nnn or PM-nnn

GelAnalysis

Initial MultiGene-12™ Product Offering

• MultiGene-12™ Human Caspase RT-PCR Profiling KitPH-001ACASP 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 13

• MultiGene-12™ Human BCL2 RT-PCR Profiling KitPH-002ABAD, BAK1, BAX, BCL2, BCL10, BCL2A1, BCL2L2, BCLxL, BCLxS, BNIP3, BIK

• MultiGene-12™ Human Integrin-I RT-PCR Profiling KitPH-003AITGA 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 2B

• MultiGene-12™ Human Integrin-II RT-PCR Profiling KitPH-004AITGA 11, ITGAL, ITGAM, ITGAV, ITGAX, ITGB1, 2, 3, 4, 5, 6

Initial MultiGene-12™ Product Offering

• MultiGene-12™ Human MMP-I RT-PCR Profiling KitPH-005AMMP 1, 2, 3, 7, 8, 9, 10, 11, 12, 13, 14

• MultiGene-12™ Human MMP-II RT-PCR Profiling KitPH-006AMMP 15, 16, 17, 20, 24, 26, MMPL1, TIMP1, 2, 3, 4

• MultiGene-12™ Human Cytokine-I RT-PCR Profiling KitPH-007AIL1B, 2, 4, 5, 6, 8, 11, 12A, 13, 15

• MultiGene-12™ Human Cytokine-II RT-PCR Profiling KitPH-008AIL16, 17, 18, 25, IFNG, IL1A, CSF2, MIF, TGFB1, 3, LTA

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

Differential Gene Expression Observed with a MultiGene-12™ RT-PCR Profiling Kit

MultiGene-12™ Human Cytokine II RT-PCR Profiling Kit

IL16

IL17

IL18

IL25

IFN

IL1A

CS

F2

MIF

TG

F1

TG

F

LTA

GA

PD

StimulatedWith PMA and A23187

Unstimulated

RNA from Jurkat cells:

RNA Interference Products:SureSilencing™ siRNA Kits

Knock down the expression of key genes in specific biological pathways

The siRNA Concept

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

How we generate siRNA Populations

PCR

T7 RNAPolymerase

RNase III

siRNA

T7 Promoter

Gene-specific cDNA fragmentsSame ones used for our GEArrays

Overview of SureSilencing™ siRNA Kit ProcedureNegative Control

siRNASureSilencing™

siRNA

LipofectAMINE2000™(Invitrogen)

transient transfection

CELLS CELLS

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.

24 to 48 h incubation

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

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

Additional Materials RequiredAccessory Products (Sold Separately):C-01: ReactionReady™ First Strand cDNA Synthesis Kit*

Enough reagents for 8 reverse transcription reactionsBuffers RI, RE, P, BC, H2O

P-1000A: ReactionReady™ PCR master mix 1000Enough lyophilized master mix for 96 reactionsEight tubes, twelve reactions each

* At this time, not available in all international territories.

Or Individual Components from Promega:Reagent ProductRandom Primers Promega Catalog # C118RNase Inhibitor Promega Catalog # N2511Reverse Transcriptase Promega Catalog # M1701dNTPs Promega Catalog # U1330Taq DNA Polymerase Promega Catalog # M2661

Initial SureSilencing™ Product Offering

• RH-0001APC Adenomatosis polyposis coli

• RH-0002YY1 YY1 transcription factor

• RH-0003E2F1 E2F transcription factor 1

• RH-0004BRCA1 Breast cancer 1, early onset

• RH-0005BAD BCL2-antagonist of cell death

• RH-0006MYC c-myc proto-oncogene

• RH-0007STAT1 Signal transducer and activator of transcription 1

• RH-0008FADD Fas-associated via death domain

• RH-0009BCL2 B-cell CLL/lymphoma 2

• RH-0010LCK 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.

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.

• NFB1 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

Relationship Between SuperArray Product Lines

GEArray™cDNA Arrays Kits

MultiGene-12™RT-PCR Profiling Kits

SureSilencing™siRNA Kits