Establishment and Verification of 6-color Fluorescent-labeled Rapid PCR Amplification System.
- Author:
Ya-ju LIU
;
Jun-tao ZHANG
;
Hai-ying JIN
;
Mei-sen SHI
- Publication Type:Journal Article
- MeSH:
Electrophoresis, Capillary;
Fluorescence;
Genetics, Population;
Genotype;
Humans;
Microsatellite Repeats;
Multiplex Polymerase Chain Reaction/methods*;
Sensitivity and Specificity
- From:
Journal of Forensic Medicine
2016;32(2):109-113
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To establish the rapid PCR amplification program and system and to verify the technical indexes.
METHODS:PCR multiplex and capillary electrophoresis detection of 24 autosomal STR loci and one Y-STR loci using the 6-color fluorescence marking technology, as well as A melogenin and Y-InDel. Meanwhile, sensitivity, specificity, identity, stability, mixing and a batch of sample tests were investigated, and the genotype of various routine samples and degraded, exfoliated cell samples were observed.
RESULTS:The sensitivity of the system was 0.062 5 ng. In addition, the genotype could be detected accurately only around 65 min via rapid amplification. The species-specificity was high and the genotyping of all kinds of dry blood specimens of filter paper and mixed, degraded, exfoliated cell samples were accurate.
CONCLUSION:The rapid amplification system can significantly improve the detection rate, and obtain accurate and stable genotyping results, which may be important implications for the establishment of STR database and study on population genetics and forensic identification.
