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Amersham CyScribe Post-Labeling Kit- For the generation of Cy3- and Cy5-labeled cDNA for microarray hybridization.
- Includes CyScribe™ reverse transcriptase.
- Produces cDNAs that are highly efficiently and evenly labeled with both Cy3 and Cy5 dyes.
- Includes ready-to-use, lyophilized aliquots of highly reactive Cy3 and Cy5 dyes.
- Optimized labeling reagents and complete protocols for labeling, purification, and quantitation of cDNA.
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Amersham CyScribe Post-Labeling Kit
Technical Information
CyScribe™ Post-Labeling Kit features a two-step protocol to prepare cDNA labeled with CyDye™ fluors*. The CyScribe reverse transcriptase synthesizes first-strand cDNA incorporating a chemically reactive nucleotide analog (aminoallyl-dUTP) into cDNA. The mRNA template is degraded and the cDNA purified to remove free nucleotides and oligomers. This cDNA is then "post labeled" with the reactive forms of Cy3- or Cy5-NHS esters, which bind to the modified nucleotides. A final purification is then performed resulting in purified CyDye-labeled cDNA, ready for hybridization. These cDNAs generate even and bright signal levels in microarray hybridization applications.
The Cy3 and Cy5 reactive dyes contain 40 000 pmol of reactive dye with greater than 75% reactive dye content. They are available in convenient individually dispensed aliquots (in amber vials) for post-labeling of cDNA. These aliquots have been sealed into foil packs to minimize degradation of the reactive groups by exposure to atmospheric humidity.
CyScribe Post-Labeling Kit includes the following reagents sufficient for 24 reactions: CyScribe reverse transcriptase, nucleotide mix, aminoallyl-dUTP, anchored oligo (dT), random nonamers, 5× CyScribe reaction buffer, 0.1 M DTT, 12 ready to use aliquots of Cy3-NHS ester and 12 ready to use aliquots of Cy5-NHS ester, nuclease-free water, control RNA: 0.24 to 9.5 kb RNA ladder, microarray hybridization buffer, and optimized protocols.
* For licensing information, see Legal Info.
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