Simple Method to Genotype the ACTN3 r577x Polymorphism
| Autor: | João Bosco Pesquero, Augusto Schneider, Ines Schadock, Márcia Rubia Duarte Buchweitz, Edgar J. Paredes-Gamero, Ronaldo C. Araujo, Carlos Castilho Barros, Marcio Nunes Corrêa, Elton Dias da Silva |
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| Rok vydání: | 2015 |
| Předmět: |
Genetics
Premature Stop Codon Base Sequence Genotyping Techniques Skeletal muscle contraction General Medicine Biology Standard methods Athletic Performance Molecular biology Polymerase Chain Reaction Polymorphism Single Nucleotide Actins law.invention law Genotype Humans Actinin Allele Restriction fragment length polymorphism Genotyping Genetics (clinical) Polymerase chain reaction DNA Primers |
| Zdroj: | Genetic testing and molecular biomarkers. 19(5) |
| ISSN: | 1945-0257 |
| Popis: | The alpha-actinin-3 r577x polymorphism (rs1815739) is one of the most important polymorphisms associated with athletic performance. This single-nucleotide mutation leads to a premature stop codon, resulting in a nonfunctional protein product. The presence of the dominant R allele is associated with full power skeletal muscle contraction. Homozygosity for the X allele is correlated with more efficient energy disposure. Restriction fragment length polymorphism and real-time polymerase chain reaction (PCR) are the standard methods used to genotype this polymorphism, but they are expensive and require special equipments. Here, we present a simple and cost-efficient method to genotype the ACTN3 r577x polymorphism by a single PCR. External primers yield a 690-bp product that indicates the template quality. Internal primers produce a 413-bp product if the R allele is present and a 318-bp product if the X allele is present. Our four-primer genotyping PCR was validated by the standard real-time PCR, generally used to genotype this single-nucleotide polymorphism, demonstrating the accuracy of this method. This protocol is perfect for small- or large-scale cohort genotyping of the ACTN3 r577x polymorphism. |
| Databáze: | OpenAIRE |
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