Asymmetric PCR is a variation of

PCR PCR or pcr may refer to: Science * Phosphocreatine, a phosphorylated creatine molecule * Principal component regression, a statistical technique Medicine * Polymerase chain reaction ** COVID-19 testing, often performed using the polymerase chain r ...

used to preferentially amplify one strand of the original DNA more than the other. The technique has applications in some types of

sequencing In genetics and biochemistry, sequencing means to determine the primary structure (sometimes incorrectly called the primary sequence) of an unbranched biopolymer. Sequencing results in a symbolic linear depiction known as a sequence which succi ...

and hybridization probing where having only one of the two complementary strands is required.

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. 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.

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. Single stranded DNA is also important for

aptamer Aptamers are short sequences of artificial DNA, RNA, XNA, or peptide that bind a specific target molecule, or family of target molecules. They exhibit a range of affinities ( KD in the pM to μM range), with little or no off-target binding ...

generation.

References

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