cDNA Generation on Paramagnetic Beads

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 (2 –4 ). This is not possible using conventional techniques of mRNA isolation and cDNA cloning.