title: Development of Novel High-Resolution Melting-Based Assays for Genotyping Two Alu Insertion Polymorphisms (FXIIIB and PV92).
abstract: 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.
authors: Department of Nutrition and Biochemistry, School of Sciences, Pontificia Universidad Javeriana, Bogotá, Colombia.|Laboratory of NeuroPsychiatric Genetics, Biomedical Sciences Research Group, School of Medicine, Universidad Antonio Nariño, Bogotá, Colombia.
authors: Yeimy
authors: Y
authors: González-Giraldo
authors: Laboratory of NeuroPsychiatric Genetics, Biomedical Sciences Research Group, School of Medicine, Universidad Antonio Nariño, Bogotá, Colombia.|Faculty of Science, Universidad Antonio Nariño, Bogotá, Colombia.
authors: Marisol
authors: M
authors: Rodríguez-Dueñas
authors: Laboratory of NeuroPsychiatric Genetics, Biomedical Sciences Research Group, School of Medicine, Universidad Antonio Nariño, Bogotá, Colombia. diego.forero@uan.edu.co.
authors: Diego A
authors: http://orcid.org/0000-0001-9175-3363
authors: DA
authors: Forero
mesh_terms: D020087:Alu Elements; D004247:DNA / Q000032:analysis; D005456:Fluorescent Dyes; D060005:Genotyping Techniques / Q000191:economics / Q000379:methods*; D006801:Humans; D054643:INDEL Mutation; D020641:Polymorphism, Single Nucleotide; D060888:Real-Time Polymerase Chain Reaction / Q000191:economics / Q000379:methods*; D015203:Reproducibility of Results; D044366:Transition Temperature
chemical_list: D005456:Fluorescent Dyes; D004247:DNA
keywords: Ancestry informative markers; Genotyping assays; High-resolution melting; Insertion/Deletion polymorphisms; Molecular genetics; Real-time PCR