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  • Title: Allele-specific PCR in SNP genotyping.
    Author: Gaudet M, Fara AG, Beritognolo I, Sabatti M.
    Journal: Methods Mol Biol; 2009; 578():415-24. PubMed ID: 19768609.
    Abstract:
    The increasing need for large-scale genotyping applications of single nucleotide polymorphisms (SNPs) in model and nonmodel organisms requires the development of low-cost technologies accessible to minimally equipped laboratories. The method presented here allows efficient discrimination of SNPs by allele-specific PCR in a single reaction with standard PCR conditions. A common reverse primer and two forward allele-specific primers with different tails amplify two allele-specific PCR products of different lengths, which are further separated by agarose gel electrophoresis. PCR specificity is improved by the introduction of a destabilizing mismatch within the 30 end of the allele-specific primers. This is a simple and inexpensive method for SNP detection that does not require PCR optimization.
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