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Mol Biotechnol ; 58(3): 197-201, 2016 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-26843017

RESUMO

Insertion/Deletion polymorphisms (InDels) are a common type of genetic variation, with a growing role in population genetics and applied genomics. There is the need for the development of novel cost-effective assays for genotyping InDels of high importance. The main objective of this study was to develop high-resolution melting-based assays for genotyping two commonly studied Alu insertion polymorphisms: FXIIIB and PV92 (rs70942849 and rs3138523). Three primers (two forward and one reverse) were designed for each marker, and high-resolution melting (HRM) analyses in a qPCR platform were performed, using EvaGreen fluorescent dye. For each one of the two Alu insertion polymorphisms, HRM analyses identified distinguishable peaks for the three genotypes, allowing a robust genotyping. Results were validated using 96 DNA samples previously genotyped and the assays worked with different DNA concentrations. In this study, we developed novel cost-effective assays, using qPCR, for genotyping two Alu insertion polymorphisms (widely used as ancestry markers). Our results highlight the feasibility of using HRM analyses for genotyping InDel polymorphisms of medical and biotechnological importance.


Assuntos
Elementos Alu , Técnicas de Genotipagem/métodos , Polimorfismo de Nucleotídeo Único , Reação em Cadeia da Polimerase em Tempo Real/métodos , DNA/análise , Corantes Fluorescentes , Técnicas de Genotipagem/economia , Humanos , Mutação INDEL , Reação em Cadeia da Polimerase em Tempo Real/economia , Reprodutibilidade dos Testes , Temperatura de Transição
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