DNA Amplification Using Promega's PowerPlex® Fusion Systems (5C and 6C).

Multiplex amplification and typing of short tandem repeat (STR) loci enable the rapid characterization of forensic samples for human identification purposes with the potential for high discriminatory power. Many commercial multiplex kits are available for consideration and purchase by DNA testing laboratories, including the PowerPlex® Fusion 5C and PowerPlex® Fusion 6C Systems offered by Promega Corporation. Herein we describe full-volume and half-volume amplification protocols utilizing extracted DNA for these respective multiplex kits, as well as procedures for direct amplification of lytic and nonlytic storage card punches and swabs.

Developmental validation of the PowerPlex(®) Fusion 6C System.

The PowerPlex(®) Fusion 6C System is a 27-locus, six-dye, multiplex that includes all markers in the expanded CODIS core loci and increases overlap with STR database standards throughout the world. Additionally, it contains two, rapidly mutating, Y-STRs and is capable of both casework and database workflows, including direct amplification. A multi-laboratory developmental validation study was performed on the PowerPlex(®) Fusion 6C System. Here, we report the results of that study which followed SWGDAM guidelines and includes data for: species specificity, sensitivity, stability, precision, reproducibility and repeatability, case-type samples, concordance, stutter, DNA mixtures, and PCR-based procedures. Where appropriate we report data from both extracted DNA samples and direct amplification samples from various substrates and collection devices. Samples from all studies were separated on both Applied Biosystems 3500 series and 6-dye capable 3130 series Genetic Analyzers and data is reported for each. Together, the data validate the design and demonstrate the performance of the PowerPlex(®) Fusion 6C System.