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• W2034649637 abstract "Allele-specific PCR (AS-PCR) has been widely used for the detection of single nucleotide polymorphism. But there are some challenges in using AS-PCR for specifically detecting DNA variations with short deletions or insertions. The challenges are associated with designing selective allele-specific primers as well as the specificity of AS-PCR in distinguishing some types of single base-pair mismatches. In order to address such problems and enhance the applicability of AS-PCR, a general primer design method was developed to create a multiple base-pair mismatch between the primer 3'-terminus and the template DNA. This approach can destabilize the primer-template complex more efficiently than does a single base-pair mismatch, and can dramatically increase the specificity of AS-PCR. As a proof-of-principle demonstration, the method of primer design was applied in colony PCR for identifying plasmid DNA deletion or insertion mutants after site-directed mutagenesis. As anticipated, multiple base-pair mismatches achieved much more specific PCR amplification than single base-pair mismatches. Therefore, with the proposed primer design method, the detection of short nucleotide deletion and insertion mutations becomes simple, accurate and more reliable." @default.
• W2034649637 created "2016-06-24" @default.
• W2034649637 creator A5051013509 @default.
• W2034649637 creator A5062667245 @default.
• W2034649637 creator A5088495278 @default.
• W2034649637 date "2010-02-01" @default.
• W2034649637 modified "2023-10-17" @default.
• W2034649637 title "Enhancing allele-specific PCR for specifically detecting short deletion and insertion DNA mutations" @default.
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• W2034649637 doi "https://doi.org/10.1016/j.mcp.2009.08.001" @default.
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• W2034649637 hasPublicationYear "2010" @default.
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