Asymmetric PCR
Asymmetric PCR is a variation of PCR used to preferentially amplify one strand of the original DNA more than the other.[1] The technique has applications in some types of sequencing and hybridization probing where having only one of the two complementary strands is required.[2]
Methodology
Asymmetric PCR differs from regular PCR by the excessive amount of primers for a 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.[3]
A modification on this process, known as Linear-After-The-Exponential-PCR (LATE-PCR), uses a limiting primer with a higher melting temperature than the excess primer to maintain reaction efficiency as the limiting primer concentration decreases mid-reaction.[4]
Applications
Asymmetric PCR can be used to form single stranded DNA from double stranded DNA, which is then used for DNA sequencing in the mutagenesis method.[citation needed] Single stranded DNA is also important for aptamer generation.[1]
References
- ^ a b Citartan, Marimuthu; et al. (December 2011). "Asymmetric PCR for good quality ssDNA generation towards DNA aptamer production" (PDF). Songklanakarin J. Sci. Technol. 34 (2): 125–131.
- ^ Wooddell, C I; Burgess, R R (1996). "Use of Asymmetric PCR to Generate Long Primers and Single-stranded DNA for Incorporating Cross-linking Analogs into Specific Sites in a DNA Probe" (PDF). Genome Res. (6): 886–892. doi:10.1101/gr.6.9.886.
- ^ Heiat, Mohammad; et al. (14 January 2017). "Essential strategies to optimize asymmetric PCR conditions as a reliable method to generate large amount of ssDNA aptamers". Biotechnology and Applied Chemistry. 64 (4): 541–548. doi:10.1002/bab.1507.
- ^ Sanchez, J. Aquiles; et al. (4 December 2003). "Linear-After-The-Exponential (LATE)–PCR: An advanced method of asymmetric PCR and its uses in quantitative real-time analysis". Proceedings of the National Academy of Sciences. 101 (7): 1933–1938. doi:10.1073/pnas.0305476101. PMC 357030.