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Synthesizing Fluorescent Target cDNA for Microarray-Based
Transcriptional Profiling: Considerations Regarding Primers and
Template RNA
Camilo Canel, Ph.D.1, and Nadine
Nassif, M.S.2
1Corning Life Sciences, 2Promega Corporation
DNA microarrays are most commonly used for determining relative transcript
abundance under two or more conditions. This typically requires the
synthesis of cDNA in the presence of modified nucleotides that may
carry, or be subsequently conjugated to, fluorescent tags having
different emission spectra. The various methods in use today differ in
terms of their labeling efficiency and yield of cDNA. The new Pronto!™
Plus Indirect System addresses the issue of differential efficiency of
Cy®3- and Cy®5-dNTP incorporation into cDNA during
reverse transcription by providing aminoallyl-modified nucleotides for
cDNA synthesis.
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