Validation of the Promega Powerplex® 16 STR Multiplex System For Forensic Casework

The Centre of Forensic Sciences, in both its Toronto and Northern Regional Laboratories (Sault Ste. Marie, Ontario), has completed internal validation of the Promega PowerPlex®16 DNA typing system in accordance with the revised SWGDAM validation guidelines (January 2004). The Promega PowerPlex®16 DNA typing system amplifies the 13 CODIS loci along with Penta D, Penta E, and Amelogenin in a single reaction, affording the laboratory greater efficiencies, discrimination power, and throughput. Methodologies, techniques and platforms already in place for current multiplex systems were employed. Detection of amplified product was achieved using 5% Long Ranger® Singel Polyacrylamide gels on ABI 377 Automated Sequencers. Robust performance of the PowerPlex®16 system was obtained using 1ng of target DNA in a 15 pL reaction amplified for a total of 30 PCR cycles. With these conditions, 0.5 pL of amplified product yielded average combined allelic peak heights in excess of 1000 rfu per locus. Sensitivity studies showed the system is capable of generating complete DNA profiles with as little as 125 pg of DNA. The PowerPlex®16 system could also consistently detect the presence of a minor component in a 19:1 mixture. Finally the system was proven to be reproducible and robust in amplifying previously typed non-probative forensic casework samples. This validation study has demonstrated that the Promega PowerPlex®16 multiplex system provides consistent amplification with a variety of different template challenges, and therefore is suitable for casework implementation in our laboratories.