Development of a new screening method to determine the main 52 mitochondrial haplogroups through a single minisequencing reaction

Leire Palencia-Madrid; Sergio Cardoso; Fernando Castro-Maestre; Igor Baroja-Careaga; Ana M Rocandio; Marian M de Pancorbo

doi:10.1016/j.mito.2018.02.004

Development of a new screening method to determine the main 52 mitochondrial haplogroups through a single minisequencing reaction

Mitochondrion. 2019 Mar:45:46-51. doi: 10.1016/j.mito.2018.02.004. Epub 2018 Feb 21.

Authors

Leire Palencia-Madrid¹, Sergio Cardoso¹, Fernando Castro-Maestre¹, Igor Baroja-Careaga¹, Ana M Rocandio², Marian M de Pancorbo³

Affiliations

¹ BIOMICs Research Group, Lascaray Research Center, University of the Basque Country (UPV/EHU), Vitoria-Gasteiz, Spain.
² Department of Nutrition and Food Sciences, Faculty of Pharmacy, University of the Basque Country (UPV/EHU), Vitoria-Gasteiz, Spain.
³ BIOMICs Research Group, Lascaray Research Center, University of the Basque Country (UPV/EHU), Vitoria-Gasteiz, Spain. Electronic address: [email protected].

PMID: 29474835
DOI: 10.1016/j.mito.2018.02.004

Abstract

This work presents the design, development and optimization of a screening method based on single-base extension sequencing to simultaneously analyze a panel of 52 mitochondrial SNPs. This enables to recognize the main mitochondrial haplogroups and to discriminate even between lineages from the same phylogenetic branch that diverged in different continents. The unavailability of individuals harboring infrequent variants was a limitation to optimize the panel. To overcome this, we have modified DNA by site-directed mutagenesis to create the unavailable allelic variants. This allowed us to verify the reliability of this panel and its usefulness to be applied in biomedicine, forensic and population genetic studies.

Keywords: Forensic genetics; Genetic epidemiology; Human population genetics; SNaPshot; Single-base extension; Site-directed mutagenesis.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Genotyping Techniques / methods*
Haplotypes*
Humans
Mass Screening / methods*
Mitochondria / genetics*
Polymorphism, Single Nucleotide
Sequence Analysis, DNA / methods*