Validation of AmpF/STR® MiniFilerTM PCR Amplification Kit for Forensic Applications

Abstract

A validation study was performed for the AmpF/STR MiniFiler PCR Amplification Kit (the MiniFiler kit) to evaluate its usefulness in analyzing forensic biological samples. The optimal DNA input for PCR amplification was approximately 0.5 ng of DNA. The marker-specific stutter ratios to filter stutter peaks provided by the kit manufacturer have been adopted without change for all markers. From a mixture of two individual DNA samples, all alleles possessed by the minor DNA contributor were detected when DNA derived from the minor contributor was present at 30% of the total DNA and above. A human genomic DNA quantitation system using real time PCR with an intercalator dye and primers targeting the 98 bp in D17Z1 was developed. The MiniFiler kit combined with the developed DNA quantitation system to determine the amount of DNA input into the PCR reaction was shown to be useful in analyzing degraded DNA samples. Primer concordance validation study using the MiniFiler and AmpF/STR Identifiler PCR Amplification Kits was performed. Typing discrepancies were observed at CSF1PO (1 individual) and D21S11 (1 individual) out of 330 Japanese individuals.