Abstract
Abstract Accurate analysis of gene expression has become achievable in recent years thanks to the development of a variety of sophisticated techniques (1). In this chapter we describe the polymerase chain reaction (PCR)-based subtractive hybridization technique of cDNA representational difference analysis (cDNA RDA). cDNA RDA can be used to identify genes whose expression differs between two or more populations, and unlike gene array approaches. requires only basic laboratory equipment. cDNA RDA is a flexible, relatively inexpensive, and highly effective technique in which target cDNA fragments are sequentially enriched by favourable hybridization kinetics and subsequently amplified by PCR. The positive selection of differences and removal of common genes targets identifi cation to only those genes which differ between populations, resulting in great sensitivity, and allowing application of the technique to very low amounts of starting material, including fluorescence-activated cell sorter (FACS) purified populations of cells.
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