The Whole Transcript (WT) Sense Target Labeling Assay Has an Updated Protocol

The GeneChip® WT (Whole Transcript) cDNA Synthesis and Amplification Kit is optimized to be used in the first steps of the GeneChip® WT Sense Target (ST) Labeling Assay.

The WT cDNA Synthesis and Amplification Kit supports the first-and second-cycle cDNA synthesis as well as the intermediate in vitro transcription amplification procedures.

The input material is 1 µg of total RNA (1 µg recommended, with an acceptible range of 1-2 µg), which is treated with the RiboMinus™ Human/Mouse Transcriptome Isolation kit (from Invitrogen, not included in this kit) to remove the majority of the ribosomal RNA for enhanced sensitivity. The resulting product from the WT cDNA Synthesis and Amplification Kit is random-primed, single-stranded DNA in the sense orientation, ready to be fragmented and labeled with the GeneChip® WT Terminal Labeling Kit (available separately from Affymetrix).

An optional method may be used to reduce the starting material to 100 ng of total RNA (100 ng recommended, with an acceptible range of 100-300ng).

Sub-Kit 1: GeneChip® WT cDNA Synthesis Kit	For 10 Rxns	For 30 Rxns
T7-(N)6 Primers, 2.5 µg/µL	20 µL	60 µL
5X 1st Strand Buffer	100 µL	300 µL
DTT, 0.1M	50 µL	150 µL
dNTP, 10 mM	9 µL	27 µL
RNase Inhibitor	25 µL	75 µL
SuperScript™ II	90 µL	270 µL
MgCl2, 1M	40 µL	120 µL
DNA Polymerase I	6 µL	18 µL
RNase H	22 µL	66 µL
Random Primers, 3 µg/µL	20 µL	60 µL
dNTP+dUTP, 10 mM	20 µL	60 µL
RNase-free Water	4.44 mL	4.44 mL

Sub-Kit 2: GeneChip® WT cDNA Amplification Kit	For 10 Rxns	For 30 Rxns
10X IVT Buffer	50 µL	150 µL
IVT NTP Mix	200 µL	600 µL
IVT Enzyme Mix	50 µL	150 µL
IVT Control	10 µL	10 µL

The WT cDNA Synthesis and Amplification Kit is also available as a component of the GeneChip® WT Sense Target Labeling and Control Reagents Kit.

Data Sheets

Exon Array System (pdf, 464 KB)

Technical Notes

Array Design for the Human Exon 1.0 ST Array (pdf, 348 KB)

Human Exon 1.0 ST Array and WT Sense Target Labeling Assay for Genome-Wide, Exon-Level Expression Analysis (pdf, 624 KB)

Application Notes

Whole-transcript Expression Analysis (pdf, 557 kb)

Part #	Description	Details
900672	GeneChip® WT cDNA Synthesis and Amplification Kit	Sufficient for 10 reactions
900673	GeneChip® WT cDNA Synthesis and Amplification Kit	Sufficient for 30 reactions
900652	GeneChip® WT Sense Target Labeling and Control Reagents	Sufficient for 30 reactions

Related Products

Part #	Description	Details
900649	GeneChip® Human Exon 1.0 ST Array	See Product Page
900817	GeneChip® Mouse Exon 1.0 ST Array	See Product Page
900820	GeneChip® Rat Exon 1.0 ST Array	See Product Page
901085	GeneChip® Human Gene 1.0 ST Array	See Product Page
901168	GeneChip® Mouse Gene 1.0 ST Array	See Product Page
901172	GeneChip® Rat Gene 1.0 ST Array	See Product Page
900720	GeneChip® Hybridization, Wash, and Stain Kit	Sufficient for 30 reactions
00-0211	GeneChip® Scanner 3000 7G with Workstation
00-0079	GeneChip® Fluidics Station 450

FAQs

Exon Array Analysis

Exon Array Design

WT Sense Target Labeling Assay

Manuals

Whole Transcript (WT) Sense Target Labeling Assay (pdf, 1.12 MB)

Package Inserts

WT cDNA Synthesis and Amplification Kit (pdf, 253 KB)

Additional Support

Fluidics Scripts

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