Amino Allyl Labeling and Hybridization Protocol

Abstract

The SuperScriptTM Plus Indirect cDNA Labeling System is a highly efficient system for generating fluorescently labeled cDNA for use on micorarrays in gene expression studies. It uses an aminoallyl-modified nucleotide together with other dNTPs in a cDNA synthesis reaction with SuperScriptTM III Reverse Transcriptase. After a purification step to remove unincorporated nucleotides, the amino-fluorescent modified cDNA is coupled with a monoreactive, Nhydroxysuccinimide (NHS)-ester fluorescent dye. A final purification step removes any unreacted dye, and the fluorescently labeled cDNA is ready for hybridization to microarrays.