A New Multiplex MiniSTR System With Three Loci And Its Application In Forensic Genetics

Objective The purpose of this project is to establish a new fluorescent multiplex miniSTR system with D19S591, D2S2944 and D18S872. Methods Potential miniSTR markers were screened from the "for the record" data in Journal of Forensic Science. PCR primers were designed for miniSTR using web-based Primer3. PCR products were analyzed with gel electrophoresis and visualized by silver staining. Using the fluorescent primers, a multiplex miniSTR system with D19S591, D2S2944 and D18S872 was constructed. The amplicons of the multiplex miniSTR system were analyzed using the ABI 310 genetic analyzer. According to the guidelines of TMGDAM in United States, we also validated the forensic application of this miniplex system. Results we successfully designed the primers for three miniSTR loci, which can generate amplicons less than 130 bp in size. The distributions of genotypes and allele frequencies of three miniSTR loci were obtained. The fluorescent miniplex system with D19S591, D2S2944 and D18S872 was successfully established. The sensitivity of this system was 0.25 ng template DNA. Its species specificity was good. After testing the aged samples and the degraded DNA mode, it was demonstrated that this miniplex system had a high success for analysis of degraded samples. Conclusions This miniplex system can be successfully used in the ABI-310 detection platform to get the reliable data. The research of forensic genetics implies that it is an easily-regulated, low cost multiplex system with stable outcome, high sensitivity, good species specificity. Based on these advantages, it could be recommended to the practice of forensic genetics. Especially it can be used in the analysis of degraded DNA specimens.