Chapter 22 - mRNA Differential Display

Abstract

The messenger RNA (mRNA) differential display process is governed by exclusive patent rights granted to GenHunter Corporation (Nashville, TN). The strategy involves coupling standard methods for complementary DNA (cDNA) synthesis to polymerase chain reaction (PCR)-based amplification using large combinations of short primers for the purpose of evaluating two or more experimental populations in a side-by-side comparison by polyacrylamide gel electrophoresis (PAGE). When identical primer sets are used, transcripts common to the RNA populations under investigation are manifested as PCR products of identical size side-by-side in adjacent lanes on a gel. This method facilitates the identification of bands in one lane that are absent or of different abundance in the corresponding location of adjacent lanes. Several variations on this standard method appear ubiquitously and are described by a variety of names. Perhaps the most scientifically correct terminology to describe this technique is mRNA differential display. However, differential display PCR is a more commonly used and instantly recognizable name. Differential display PCR is all about the speed, specificity, and sensitivity of PCR, by which at least a section of each of the various transcripts being produced by the cell is amplified in the form of cDNA. The strategy behind differential display PCR (DDPCR) is simple: incorporation of traditional synthesis of cDNA from purified RNA followed by an amplification of the products of the first-strand cDNA reaction. Among the advantages of the DDPCR method is the requirement for small input RNA mass.