Validation of Half-Reaction Amplification Using Promega PowerPlex® 16
DNA amplification is a fundamental yet costly process used in DNA analysis. This study evaluated half-reaction amplification (12.5, 12, and 13 uL) using the Promega Powerplex® 16 Kit with the hope of reducing sample analysis costs by half. A sensitivity study was completed, along with the testing of various blood stain samples including those with low (<0.40 ng) and high DNA concentrations (>3.0 ng), peak height imbalances, and allelic drop-out. Also, 467 samples submitted to the MUFSC laboratory for testing were analyzed. Results indicate that half-reaction amplification produced higher quality profiles than full-reactions. Average peak heights increased by 85%, peak height imbalances improved, and drop-out was eliminated in 75.8% of samples. Only eight of 467 case samples required re-amplification, a success rate of 94% was observed, and the repeat rate decreased significantly. Finally, a DNA input of 0.25–1.0 ng is ideal for half-reaction amplification.
Keywords: CSF; D13S317; D16S539; D18S51; D21S11; D3S51358; D5S818; D7S820; D8S1179; DNA amplification; FGA; Penta D; Penta E; PowerPlex® 16; THO1; TPOX; forensic DNA; forensic science; vWA; validation
Document Type: Research Article
Affiliations: Marshall University Forensic Science Center, DNA Laboratory, 1401 Forensic Science Drive, Huntington, WV.
Publication date: July 1, 2010