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

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Contains All Reagents Required for Double-Stranded cDNA Synthesis Within the Affymetrix® GeneChip® Workflow

Now available from Invitrogen, 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 Invitrogen’s 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 (Table 1)
  • Reduces costs and insures supply because you buy directly from the manufacturer Perform more full-length double-stranded cDNA synthesis within the Affymetrix® One-Cycle workflow using a qualified T7-Oligo(dT) Promoter Primer and SuperScript® II Reverse Transcriptase, a proprietary RNase H–reduced RT only available from Invitrogen (Figure I).

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 (Invitrogen) and the GeneChip® One-Cycle cDNA Synthesis Kit (Affymetrix), cRNA yields were comparable (Figure 2). In addition, both the SuperScript® One-Cycle cDNA Kit (Invitrogen) and the GeneChip® One-Cycle cDNA Synthesis Kit (Affymetrix) delivered fully representative cDNA outputs as indicated by the equivalent 3´ to 5´ ratios for the GAPDH probe set (Figure 3).

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.

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