chapter  20
4 Pages

Differential Screening by RDA: Representational Difference Analysis

The subtraction of cDNA by RDA is a differential screening technique assisted by PCR, which allows the identification of genes expressed specifically in a target tissue in relation to a control tissue. The subtraction comprises the following steps: (1) synthesis of double-stranded cDNAs and their cleavage by a restriction enzyme; (2) their multiplication by PCR amplification; (3) subtractive hybridization. Several cycles of subtractive hybridization are effected so as to enrich the product obtained in differentially expressed genes.