Asymmetric PCR
Asymmetric PCR Protocol
Asymmetric PCR is used to preferentially amplify one strand of the target DNA more than the other.
Applications of Asymmetric PCR
The Asymmetric PCR is useful in some sequencing and hybridization probing applications where having only one of the two complementary stands is sufficient or required.
Asymmetric PCR Method
The asymmetric PCR method is conducted as the standard pcr protocol, however a great excess of the primers for the chosen strand is used.
Due to the slow (arithmetic) amplification later in the reaction after the limiting primer has been used up, extra cycles of PCR are required.
Also see Linear After the Exponential PCR or LATE-PCR.
Primers for Asymmetric PCR
A PCR in which the predominant product is a single-stranded DNA, as a result of unequal primer concentrations.
As asymmetric PCR proceeds, the lower concentration primer is quantitatively incorporated into double-stranded DNA. The higher concentration primer continues to primer synthesis, but only of its strand.
Troubleshooting Asymmetric PCR
Asymmetric PCR References
Saiki RK, Bugawan TL, Horn GT, Mullis KB, Erlich HA (1986). "Analysis of enzymatically amplified beta-globin and HLA-DQ alpha DNA with allele-specific oligonucleotide probes.". Nature 324: 163-166. PMID 3785382.