An Updated Validation of Promega’s PowerPlex® 16 System: High Throughput Databasing Under Reduced PCR Volume Conditions on Applied Biosystem’s 96 Capillary 3730xl DNA Analyzer
The PowerPlex® 16 System from Promega Corporation allows single tube multiplex amplification of sixteen short tandem repeat (STR) loci including all 13 core combined DNA index system STRs. This report presents an updated validation of the PowerPlex® 16 System on Applied Biosystem’s 96 capillary 3730xl DNA Analyzer. The validation protocol developed in our laboratory allows for the analysis of 1536 loci (96 × 16) in c. 50 min. We have further optimized the assay by decreasing the reaction volume to one-quarter that recommended by the manufacturer thereby substantially reducing the total cost per sample without compromising reproducibility or specificity. This reduction in reaction volume has the ancillary benefit of dramatically increasing the sensitivity of the assay allowing for accurate analysis of lower quantities of DNA. Due to its substantially increased throughput capability, this extended validation of the PowerPlex® 16 System should be useful in reducing the backlog of unanalyzed DNA samples currently facing public DNA forensic laboratories.