Establishment and Verification of 6-color Fluorescent-labeled Rapid PCR Amplification System
LIU YA-JU1, ZHANG JUN-TAO2, JIN HAI-YING3, SHI MEI-SEN4
（1. Institute of Criminal Science and Technology, Xuchang Public Security Bureau, Xuchang 461000, China; 2. Criminal Investigation Team, Xiangcheng Public Security Bureau, Xiangcheng 461700, China; 3. HEALTH Gene Technologies Incorporation, Ningbo 315000, China; 4. Key Laboratory of Evidence Science, Ministry of Education, China University of Political Science and Law, Beijing 100088, China）
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 Amelogenin 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.