Validation of the DNATyper~(TM)15 PCR amplification kit / 中国法医学杂志

ABSTRACT

Objective Developmental validation studies were designed according to the standards of forensic DNA community on DNATyperTM15 kit,which simultaneously amplifies 14 STR loci(D6S1043、D21S11、D7S820、CSF1PO、D2S1338、D3S1358、D13S317、D8S1179、D16S539、Penta E、D5S818、VWA、D18S51、FGA)and amelogenin,a sex-determining locus.Methods Several key factors were tested includingⅠ.amount of hot-start Taq polymerase;Ⅱ.annealing temperature;Ⅲ.sensitivity;Ⅳ.reaction volume;Ⅴ.cycle number;Ⅵ.primer concentration.DNATyperTM15 was also compared with two other widely used commercial STR amplification kits,namely IdentifilerTM and PowerPlex.Results No difference in performance was observed with three lots of DNATyperTM15.Performance was not affected even after 20 times of repeated freeze-and-thaw.All three kits performed comparably in the following aspectsⅠ.sensitivity;Ⅱ.ability to genotype mixed samples;Ⅲ.amplification of DNA from various sample sources.DNA extraction methods(Chelex-100,magnetic beads,silica beads)did not result in any observable effect on performance with any of the three kits.Conclusion All the results demonstrated that DNATyperTM15 is suitable for both forensic DNA database work and casework.