Uses Reactive CyDye fluorescent dyes: optimized for microarray probe labeling Provides conveniently packaged, highly reactive Cy3 and Cy5 monofunctional NHS ester: to label probes via the postlabeling (amino allyl) route for use in microarray applications In the first step, reverse transcriptase synthesizes firststrand cDNA, incorporating a chemically reactive nucleotide analog (amino allyldUTP) into the cDNA. The mRNA template is then degraded and the cDNA purified to remove free nucleotides and primers. In the second step, the purified cDNA is postlabeled with the reactive forms of Cy3 or Cy5 NHS esters that bind to the modified amino allylnucleotides. After a final purification, the labeled cDNA is ready for hybridization.
