Indirect Aminoallyl-dUTP Labeling of RNA for Microarrays

Abstract

The labeling procedure described here is called indirect because the fluorescent moiety is not incorporated during the reverse transcription reaction. Instead, a reactive nucleotide analog (aminoallyl-dUTP) is incorporated during reverse transcription, the cDNA is isolated, and then the cyanine dyes are incorporated by binding with the aminoallyl group to produce the desired fluorescent cDNA.