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Amersham CyScribe First-Strand cDNA Labeling Kit- For the generation of Cy3- and Cy5-labeled cDNA for microarray hybridization analysis.
- Includes CyScribe™ reverse transcriptase.
- Optimized labeling protocol resulting in excellent incorporation of both Cy3- and Cy5-labeled nucleotides.
- Optimized reagents for use with Cy3- and Cy5-labeled dCTP or dUTP.
- Complete protocols for labeling, purification, and quantitation of cDNA.
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Amersham CyScribe First-Strand cDNA Labeling Kit
Technical Information
CyScribe™ First-Strand cDNA Labeling Kit uses CyScribe reverse transcriptase to synthesize Cy3- and Cy5-labeled first-strand cDNAs for use in microarray applications. From 50 ng to 1 μg of mRNA (or 5 to 25 μg total RNA) can be labeled in a standard 20 μl reaction in less than 3 h. The CyScribe Kit has been designed to offer the flexibility of performing labeling reactions with either Cy3- and Cy5-labeled dCTP or Cy3- and Cy5-labeled dUTP. The labeling reactions can be primed with anchored oligo(dT), which provides a convenient means for directing the synthesis of cDNA from the 3’-end of the mRNA, and with random nonamers, which will prime cDNA synthesis along the length of transcripts, or a combination of both.
Equal amounts of two different purified first-strand cDNAs labeled with Cy3 and Cy5 were hybridized with a microarray slide of spotted human cDNA clones.
CyScribe First-Strand cDNA Labeling Kit includes the following reagents sufficient for 25 labeling reactions: anchored oligo(dT), random nonamers, 5× CyScribe buffer, 0.1 M DTT, nucleotide mix for dUTP, nucleotide mix for dCTP, CyScribe reverse transcriptase, nuclease-free water, control RNA: 0.24 to 9.5 kb RNA ladder, microarray hybridization buffer and protocol booklet.
Additionally, the complete systems contain Cy3- and Cy5-labeled nucleotides* of either dCTP or dUTP.
* For licensing information, see Legal Info.
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