Fluorescent sample labeling for DNA microarray analyses

Abstract

Three fluorophor-labeling methods for gene expression profiling on deoxyribonucleic acid (DNA) microarrays are described. All three techniques start from total ribonucleic acid (RNA) samples. Two procedures are based on first-strand complementary DNA synthesis by reverse transcription. Label is introduced either by direct incorporation of fluorescently labeled nucleotides or indirectly by incorporation of aminoally-dUTP and subsequent coupling of fluorescent dyes. The third method is based on an amplification of antisense RNA by in vitro transcription subsequent to first- and second-strand complementary DNA synthesis. While the first two methods are applied mainly in analyses on microarrays made from spotted polymerase chain reaction products or long oligonucleotides, the last procedure is mostly used for experiments on in situ synthesized oligonucleotide arrays.