Figure 2

Schematic diagram of the Suppression Subtractive Hybridization protocol. Source: Clontech manual modified. Tester cDNA was divided into two aliquots, one for ligation with adapter A and the second with adapter B. Each half was then hybridized with driver cDNA that was not ligated to any adapter. Single-stranded (non-hybridized) sequences of the two independent hybridization steps were then mixed in a second hybridization step generating A-B hynrids. Primers to these adapters were then used for the selective amplification of these hybrids by PCR. This procedure allows for the enrichment of the cDNA population that is overrepresented (differentially represented) in the respective tester cDNA sample.