Figure 1
From: SuperSelective primer pairs for sensitive detection of rare somatic mutations
Design of PCR assays that utilize SuperSelective primer pairs for the selective amplification of rare mutant DNA target fragments in the presence of abundant DNA fragments from the entire human genome. After the SuperSelective primer pair initiates synthesis of amplicons on mutant target fragments (but does not initiate amplification on closely related wild-type DNA fragments), the resulting amplicons contain each primer’s bridge sequence in place of their target’s intervening sequence complement. Consequently, subsequent exponential amplification is highly efficient, since the entire sequence of each SuperSelective primer is complementary to the amplicons. Since the concentration of the forward primer is limited, and is eventually used up, the excess reverse primer continues linear amplification, creating readily available targets for molecular beacon probes, whose fluorescent color identifies their target amplicons.
