| Chapter title |
Allele-specific PCR in SNP genotyping.
|
|---|---|
| Chapter number | 26 |
| Book title |
Single Nucleotide Polymorphisms
|
| Published in |
Methods in molecular biology, September 2009
|
| DOI | 10.1007/978-1-60327-411-1_26 |
| Pubmed ID | |
| Book ISBNs |
978-1-60327-410-4, 978-1-60327-411-1
|
| Authors |
Gaudet M, Fara AG, Beritognolo I, Sabatti M, Muriel Gaudet, Anna-Giulia Fara, Isacco Beritognolo, Maurizio Sabatti, Gaudet, Muriel, Fara, Anna-Giulia, Beritognolo, Isacco, Sabatti, Maurizio |
| 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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Mendeley readers
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|---|---|---|
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| Researcher | 32 | 15% |
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| Student > Postgraduate | 10 | 5% |
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