title: [AS-PCR assay for 20 mtDNA SNP typing and haplotype frequency].
abstract: OBJECTIVE
  To develop a multiplex allele-specific PCR (AS-PCR) assay with three-color fluorescence labeling for mitochondrial DNA (mtDNA) SNP typing.
  
  
  METHODS
  Based on the principle of AS-PCR, the primer sets were designed for 20 SNP located on the coding region of mtDNA and divided into 2 groups labeled with FAM and HEX fluorescence, respectively. A primer set included two forward (reverse) allelic specific primers with different sizes and a generic reverse (forward) primer. Blood samples from 200 unrelated individuals were analyzed by AS-PCR and capillary electrophoresis. Three random samples at least for each SNP site were examined and verified by direct sequencing. The haplotype frequency was investigated.
  
  
  RESULTS
  Distinct electropherograms of 200 blood samples were obtained successfully. The typing results of direct sequencing were identical to those obtained from AS-PCR. The minimum detectable DNA concentration was 0.2 pg under the system of 10 microL. The sensitivity of the DNA concentrations ranged from 0.5 to 5 pg. The 200 individuals were assigned into 15 haplotype, and the haplotype diversity was 0.906 0.
  
  
  CONCLUSIONS
  AS-PCR is a simple, rapid and efficient method for mtDNA SNP typing, and can be applied to forensic practice.
authors: Yan-Chai
authors: YC
authors: Nie
authors: Chen
authors: C
authors: Zhang
authors: Ya-Nan
authors: YN
authors: Liu
authors: Jiang-Ping
authors: JP
authors: Huang
authors: Hai-Tao
authors: HT
authors: Jiao
authors: Dan
authors: D
authors: Wu
authors: Huai-Gu
authors: HG
authors: Zhou
mesh_terms: D000483:Alleles; D004247:DNA; D017931:DNA Primers; D004272:DNA, Mitochondrial / Q000032:analysis*; D019075:Electrophoresis, Capillary; D006239:Haplotypes; D006801:Humans; D008928:Mitochondria; D016133:Polymerase Chain Reaction / Q000379:methods*; D020641:Polymorphism, Single Nucleotide; D017422:Sequence Analysis, DNA
chemical_list: D017931:DNA Primers; D004272:DNA, Mitochondrial; D004247:DNA