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High‐Resolution Melting (HRM) of Hypervariable Mitochondrial DNA Regions for Forensic Science
Authors:Alípio dos Santos Rocha M.Sc.  Isis Salviano Soares de Amorim M.Sc.  Tatiana de Almeida Simão Ph.D.  Adenilson de Souza da Fonseca Ph.D.  Rodrigo Grazinoli Garrido Ph.D.  Andre Luiz Mencalha Ph.D.
Affiliation:1. Departamento de Biofísica e Biometria, Instituto de Biologia Roberto Alcantara Gomes, Universidade do Estado do Rio de Janeiro (UERJ), Rio de Janeiro, RJ, Brazil;2. Polícia Civil do Estado do Rio de Janeiro, Instituto de Pesquisas e Perícias em Genética Forense—IPPGF, Rio de Janeiro, RJ, Brazil;3. Departamento de Bioquímica, Instituto de Biologia Roberto Alcantara Gomes, Universidade do Estado do Rio de Janeiro (UERJ), Rio de Janeiro, RJ, Brazil
Abstract:Forensic strategies commonly are proceeding by analysis of short tandem repeats (STRs); however, new additional strategies have been proposed for forensic science. Thus, this article standardized the high‐resolution melting (HRM) of DNA for forensic analyzes. For HRM, mitochondrial DNA (mtDNA) from eight individuals were extracted from mucosa swabs by DNAzol reagent, samples were amplified by PCR and submitted to HRM analysis to identify differences in hypervariable (HV) regions I and II. To confirm HRM, all PCR products were DNA sequencing. The data suggest that is possible discriminate DNA from different samples by HRM curves. Also, uncommon dual‐dissociation was identified in a single PCR product, increasing HRM analyzes by evaluation of melting peaks. Thus, HRM is accurate and useful to screening small differences in HVI and HVII regions from mtDNA and increase the efficiency of laboratory routines based on forensic genetics.
Keywords:forensic science  high‐resolution melting  hypervariable regions  mitochondrial     DNA     melting curve
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