SuperScript® One-Cycle cDNA Kit
Maintain consistency without downtime in your Affymetrix GeneChip® workflow SuperScript® One-Cycle cDNA Kit for use with Affymetrix® One-Cycle Assays
Contains All Reagents Required for Double-Stranded cDNA Synthesis Within the Affymetrix® GeneChip® Workflow
Designed to consistently produce higher yields of full-length cDNA, the SuperScript® One-Cycle cDNA Kit for use with Affymetrix® One-Cycle Assays conveniently contains all of the reagents required for double-stranded cDNA synthesis within the Affymetrix® GeneChip® workflow.
- Contains our proprietary RNase H–reduced SuperScript® II Reverse Transcriptase for higher yields of full-length cDNA
- Includes all of the components required to carry out double-stranded cDNA synthesis, including a qualified T7-oligo(dT) promoter primer
- Reduces costs and insures supply because you buy directly from the manufacturer
After obtaining your cDNA product, you can then proceed directly to the Affymetrix® protocol to perform purification and in vitro transcription and subsequent hybridization of biotin-labeled cRNA to Affymetrix GeneChip® arrays.
Achieve Maximum Yields of Full-Length Product
When equivalent amounts of HeLa total RNA were processed using the SuperScript® One-Cycle cDNA Kit (Life Technologies) and the GeneChip® One-Cycle cDNA Synthesis Kit (Affymetrix), cRNA yields were comparable (Figure 1). In addition, both the SuperScript® One-Cycle cDNA Kit and the GeneChip® One-Cycle cDNA Synthesis Kit delivered fully representative cDNA outputs as indicated by the equivalent 3´ to 5´ ratios for the GAPDH probe set (Figure 2).
Figure 1. 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 2. 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.