A single multiplex PCR and SNaPshot minisequencing reaction of 42 SNPs to classify admixture populations into mitochondrial DNA haplogroups

Greiciane G Paneto; Stephan Köhnemann; Joyce A Martins; Regina M B Cicarelli; Heidi Pfeiffer

doi:10.1016/j.mito.2010.12.010

A single multiplex PCR and SNaPshot minisequencing reaction of 42 SNPs to classify admixture populations into mitochondrial DNA haplogroups

Mitochondrion. 2011 Mar;11(2):296-302. doi: 10.1016/j.mito.2010.12.010. Epub 2010 Dec 21.

Authors

Greiciane G Paneto¹, Stephan Köhnemann, Joyce A Martins, Regina M B Cicarelli, Heidi Pfeiffer

Affiliation

¹ Faculdade de Ciências Farmacêuticas, Unesp-Univ Estadual Paulista, Araraquara, SP, Brazil. greiciane_gp@hotmail.com

PMID: 21172459
DOI: 10.1016/j.mito.2010.12.010

Abstract

SNaPshot minisequencing reaction is in increasing use because of its fast detection of many polymorphisms in a single assay. In this work we described a highly sensitive single nucleotide polymorphisms (SNPs) typing method with detection of 42 mitochondrial DNA (mtDNA) SNPs in a single PCR and SNaPshot multiplex reaction, in order to allow haplogroup classification in Latin American admixture population. We validated the panel typing 160 Brazilian individuals. Complete SNP profiles were obtained from 10 pg of total DNA. We conclude that it is possible to build and genotype more than forty mtDNA SNPs in a single multiplex PCR and SNaPshot reaction, with sensitivity and reliability, resolving haplogroup classification in admixture populations.

Publication types

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

MeSH terms

Base Sequence
Brazil
DNA Primers
DNA, Mitochondrial / genetics*
Forensic Genetics
Haplotypes*
Humans
Polymerase Chain Reaction / methods*
Polymorphism, Single Nucleotide*
Reproducibility of Results

Substances

DNA Primers
DNA, Mitochondrial