Testing MiniSTR Primers for Addition to a PCR-Based Forensic Specimen Identification Protocol

An assay has been developed for polymerase chain reaction (PCR)-based human identity testing using the Federal Bureau of Investigation's human Combined DNA Identity System (CODIS) primers. Recent forensic literature has identified difficulties using these primers due to amplicon size and the degraded nature of DNA from forensic samples. Primers termed mini Short-Tandem Repeat (STR) primers targeted to the same loci as the CODIS primers but which have smaller amplicons have been developed. Two of the three miniSTR primer pairs examined with our established assay were successfully tested with forensic DNA. This allowed the substitution of new primers for detection of a locus, D16S539, which was poorly defined in our assay and the addition of a primer pair for a locus, FGA, not previously included. The replacement of the D16S539 CODIS primers with the miniSTR primers will provide more accurate results for this locus. The addition of the FGA mini STR primers to the core set of tested loci will increase the overall power of discrimination of our assay. Work was accomplished under approved task AM-TOXLAB.