SuperScript® One-Cycle cDNA Kit
Contains all of the reagents required for double-stranded cDNA synhesis within Affymetrix® GeneChip® workflow
|T7 Oligo(dT), 50 uM||60|
|5X 1st Strand Rxn Mix||120|
|dNTP, 10 mM||120|
|5X 2nd Strand Rxn Mix||900|
|E.coli DNA Ligase||30|
|T4 DNA Polymerase||60|
|E.coli DNA Polymerase||120|
Achieve maximum yields of full-length product
Figure 2—SuperScript® One-Cycle cDNA Kit delivers consistency in yields. Equal amounts of total HeLa RNA (either 1 μg or 15 μg) were incubated at 42°C for one hour with SuperScript® II Reverse Transcriptase (included in both kits) in the presence of T7-oligo(dT) primer for cDNA synthesis, and followed up with RNase H–mediated second-strand synthesis. Equivalent amounts of cDNA from each reaction were then processed with the Sample Cleanup Module and IVT Labeling Kit for cRNA synthesis (Affymetrix) under standard conditions. Equivalent cRNA yields are obtained from the the Affymetrix® and Invitrogen cDNA Kits under standard conditions.
Figure 3—SuperScript® One-Cycle cDNA Kit delivers fully representative cDNA product. Arrays were hybridized using 1 μg and 15 μg cDNA in a GeneChip® Hybridization Oven (Affymetrix) at 45˚C for 16 hours at 60 rpm, followed by washing in GeneChip® Fluidics Station 450 (Affymetrix, using Protocol EukGEWS2v5_450). The arrays were scanned with GeneChip® Scanner 3000 7G (Affymetrix) for 532 nm fluorescence signal. Signal on individual features was determined using GeneChip® Operating Software. Equivalent GAPDH signal ratios are obtained from the Affymetrix® and Invitrogen cDNA Kits under standard conditions.