chapter  18
4 Pages

Differential Display RT-PCR

The technique of differential display RT-PCR is a variant of differential screening adapted to the identification of rare mRNA (see Profile 17). It consists of three major steps: (1) reverse transcription from mRNAs resulting in the obtaining of single-stranded cDNAs (see Profile 27); (2) PCR amplification of the cDNAs; (3) electrophoretic separation of fragments of double-stranded cDNAs before they are cloned. These three steps are carried out simultaneously and in parallel on mRNAs isolated from different tissues or treatments. The electrophoretic gels can be compared to identify bands of cDNA that are differentially accumulated.