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Detection of 13 Hypervariable Region 1 (Hv1) SNPs using single-base extension (Sbe) primers in parallel with Sanger sequencing.
Konjhodzic, Rijad; Salihefendic, Lana; Ceko, Ivana; Kandic, Enis; Asic, Adna; Kubat, Milovan.
Afiliação
  • Konjhodzic R; Alea Genetic Center, Olovska, 67, 71000 Sarajevo, Bosnia and Herzegovina.
  • Salihefendic L; Alea Genetic Center, Olovska, 67, 71000 Sarajevo, Bosnia and Herzegovina.
  • Ceko I; Alea Genetic Center, Olovska, 67, 71000 Sarajevo, Bosnia and Herzegovina; Department of Genetics and Bioengineering, International Burch University, Francuske revolucije bb, 71000 Sarajevo, Bosnia and Herzegovina.
  • Kandic E; Health Institute "Alea Dr. Kandic", Avdage Sahinagica, 3, 71000 Sarajevo, Bosnia and Herzegovina.
  • Asic A; Department of Genetics and Bioengineering, International Burch University, Francuske revolucije bb, 71000 Sarajevo, Bosnia and Herzegovina. Electronic address: adna.asic@ibu.edu.ba.
  • Kubat M; Institute for Forensic Medicine, Medical Faculty, University of Zagreb, Salata, 11, 10000 Zagreb, Croatia.
Gene ; 872: 147438, 2023 Jul 01.
Article em En | MEDLINE | ID: mdl-37084986
The aim of this study was to determine whether single-base extension (SBE) chemistry can be applied to forensic practice of testing the target single nucleotide polymorphisms (SNPs) of the mitochondrial DNA (mtDNA) Hypervariable Region 1 (HV1). Despite itsweak discrimination power, high copy number of mtDNA per cell and its stability against degradation guarantee mtDNA testing a place in modern forensic genetics. In this research, buccal swab samples were obtained from 294 individuals from Bosnia and Herzegovina. Following DNA isolation using QIAamp® DNA Mini Kit, full sequencing of HV1 was completed using chain-termination Sanger sequencing method. SBE reactions were then performed by targeting 13 SNPs that were identified to be the most frequent in the study population. Uniplex SBE reactions for each individual SNP, as well as two multiplex reactions were prepared for both pure and mixed samples. The results showed complete agreement of the Sanger sequencing results with SBE reactions for both uniplex and multiplex reactions. The results obtained with SBE were encouraging in regard to multiplexing and processing of the mixed samples, since the allele of the minor contributor to the sample was observed in SBE electropherogram in all prepared mixtures. SBE method is limited by the fact that only target SNPs of interest will be analyzed, meaning that they must be carefully selected and curated for each population. However, typing with SBE protocol is accurate, as compared to the golden standard of Sanger sequencing, but was more time- and labor-efficient and simpler to analyze, therefore offering a suitable alternative when Sanger sequencing is not available, given that polymorphic SNPs are known for the population of interest.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: DNA Mitocondrial / Polimorfismo de Nucleotídeo Único Tipo de estudo: Diagnostic_studies / Guideline Limite: Humans Idioma: En Revista: Gene Ano de publicação: 2023 Tipo de documento: Article

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: DNA Mitocondrial / Polimorfismo de Nucleotídeo Único Tipo de estudo: Diagnostic_studies / Guideline Limite: Humans Idioma: En Revista: Gene Ano de publicação: 2023 Tipo de documento: Article