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 3′ 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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© 2009 Humana Press, a part of Springer Science+Business Media, LLC 2003
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Gaudet, M., Fara, AG., Beritognolo, I., Sabatti, M. (2009). Allele-Specific PCR in SNP Genotyping. In: Komar, A. (eds) Single Nucleotide Polymorphisms. Methods in Molecular Biology™, vol 578. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60327-411-1_26
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DOI: https://doi.org/10.1007/978-1-60327-411-1_26
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