AS-PCR assay for 20 mtDNA SNP typing and haplotype frequency / 法医学杂志
Journal of Forensic Medicine
; (6): 96-109, 2014.
Article
de Zh
| WPRIM
| ID: wpr-983888
Bibliothèque responsable:
WPRO
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.@*CONCLUSION@#AS-PCR is a simple, rapid and efficient method for mtDNA SNP typing, and can be applied to forensic practice.
Texte intégral:
1
Indice:
WPRIM
Sujet Principal:
Haplotypes
/
ADN
/
ADN mitochondrial
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Réaction de polymérisation en chaîne
/
Analyse de séquence d'ADN
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Amorces ADN
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Électrophorèse capillaire
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Polymorphisme de nucléotide simple
/
Allèles
/
Mitochondries
Limites du sujet:
Humans
langue:
Zh
Texte intégral:
Journal of Forensic Medicine
Année:
2014
Type:
Article