SuperScript One-Cycle cDNA Kit
Maintain consistency without downtime in your Affymetrix™ GeneChip™ workflow. Invitrogen™ SuperScript™ One-Cycle cDNA Kit for use with Affymetrix One-Cycle Assays.
Contains all of the reagents required for double-stranded cDNA synthesis within 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 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 Affymetrix 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.