Asymmetric PCR

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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

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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

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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

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