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3. cDNA Generation on Paramagnetic Beads
Abstract
Synthesis of complementary DNA (cDNA) by reverse transcription (RT) is a key step in investigating specific gene expression of a single transcript by RT-PCR (polymerase chain reaction) or to study the more complex profiles of gene expression in a biological sample using cDNA library or other techniques. Solid-phase generation of cDNA (1), based on immobilization of the synthesized cDNA on paramagnetic beads and the magnetic-bead separation technology, is a very useful method for studying gene expression particularly when handling limited amounts of starting materials (see Note 1). Using this method, analysis of gene expression can be carried out at a single-cell level (24). This is not possible using conventional techniques of mRNA isolation and cDNA cloning.
Affiliation(s): (2) Western Australia State Agricultural Biotechnology Center, Murdoch University, Perth, WA
Series: Methods in Molecular Biology  |  Volume: 221  |  Pub. Date: Feb-19-2003  |  Page Range: 25-31  |  DOI: 10.1385/1-59259-359-3:25
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