Differential Display: a Novel Pcr-based Method for Gene Isolation and Cloning1

Differential Display (DDRT-PCR) is a powerful technique for analyzing differences in gene expression. DDRT-PCR has the following essential steps: total RNA isolation and purification, cDNA synthesis from mRNAs, PCR amplification of cDNAs, visualization of PCR products, reamplification and cloning of the differentially expressed PCR products, confirmation by northern blot, sequencing the confirmed clones, and finally cDNA library screening to isolate the genes of interest. After its invention in 1992, a number of modifications have been introduced to optimize the technique and specifically to reduce the major problem of “false positives”. Since understanding of specific gene expression patterns that regulate developmental and stress responses is a major concern of molecular biology, DDRT-PCR has become a very popular molecular technique during the past decade. Additional