Differential Display

Abstract

Differential display has been developed as a tool to detect and characterize altered gene expression in eukaryotic cells. In this technique, as first described by Liang and Pardee (1992) (1,2) total RNA is isolated from two (or more) cell types or tissues to be compared. First strand copies of both RNAs are made by reverse transcription, using an oligo-dT primer (the anchor primer) that has a specific dinucleotide at its 3′ end (see Table 1). This anchor primer and a random 10-mer (arbitrary primer) (Table 1) are then used to amplify, by polymerase chain reaction (PCR), cDNAs to which the 3′-anchor and 5′-arbitrary primers both hybridize (Fig. 1). A radioactive nucleotide is included in the PCR reactions so that the PCR products can be run side-by-side on a 5% polyacrylamide gel and visualized by autoradiography.

Table 1 Oligonucleotide Primers Used in Differential Display