title: Multiplex mutagenically separated polymerase chain reaction assay for rapid detection of human mitochondrial DNA variations in coding area.
abstract: Recently, it has been recognized that information in the mitochondrial DNA (mtDNA) coding region can provide additional forensic discrimination with respect to the standard typing of the D-loop region, increasing the forensic power of mtDNA testing, which is sometimes rather limited. In the present study, we simultaneously typed ten single nucleotide polymorphisms (SNP) in the coding region by use of mutagenically separated polymerase chain reaction (MS-PCR) in the Chinese Chengdu population. This technique, in which different-size allele-specific primers were used, specifically amplified both alleles of mtDNA in the same reaction. Subsequent gel electrophoresis showed ten of the allelic products of different loci. Using multiplex MS-PCR, 30 primers were added simultaneously into one reaction tube to identify ten SNPs. The mtDNA variations of 160 individuals from the Chinese Chengdu population were examined and classified into 18 haplotypes. The multiplex MS-PCR method is suitable for large-scale screening studies of mtDNA variability because it is both rapid and economical.
authors.affiliation: 1Institute of Forensic Medicine, Sichuan University, Chengdu, Sichuan, People's Republic of China. wujin2028@gmail.com
authors.forename: Jin
authors.initials: J
authors.lastname: Wu
authors.forename: Peng
authors.initials: P
authors.lastname: Ran
authors.forename: Beilei
authors.initials: B
authors.lastname: Zhang
authors.forename: Yingbi
authors.initials: Y
authors.lastname: Li
authors.forename: Jing
authors.initials: J
authors.lastname: Yan
authors.forename: Miao
authors.initials: M
authors.lastname: Liao
authors.forename: Yiping
authors.initials: Y
authors.lastname: Hou
authors.forename: Lin
authors.initials: L
authors.lastname: Zhang
mesh_terms: D000483:Alleles; D001483:Base Sequence; D001681:Biological Assay; D002681:China; D004272:DNA, Mitochondrial / Q000032:analysis*; D005784:Gene Amplification; D005820:Genetic Testing; D014644:Genetic Variation; D005838:Genotype; D006801:Humans; D066298:In Vitro Techniques; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic; D020641:Polymorphism, Single Nucleotide; D013997:Time Factors
chemical_list: D004272:DNA, Mitochondrial