Asymmetric PCR

Asymmetric PCR is used to preferentially amplify one strand of the original DNA more than the other.^[1] It finds use in some types of sequencing and hybridization probing where having only one of the two complementary strands is required. PCR is carried out as usual, but with a great excess of the primers for the chosen strand. Due to the slow (arithmetic) amplification later in the reaction after the limiting primer has been used up extra cycles of PCR are required. A recent modification on this process, known as Linear-After-The-Exponential-PCR (LATE-PCR), uses a limiting primer with a higher melting temperature (Melting temperature/Tm) than the excess primer to maintain reaction efficiency as the limiting primer concentration decreases mid-reaction.

Applications

Formation of single stranded DNA from double stranded DNA which is used for DNA sequencing in mutagenesis method.

References