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 microL) 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.
Hoffman
Nicole H
NH
Marshall University Forensic Science Center, DNA Laboratory, 1401 Forensic Science Drive, Huntington, WV, USA. Nhoffman2@isp.in.gov
Fenger
Terry
T
D000483:Alleles; D004247:DNA / Q000032:analysis* / Q000235:genetics; D016172:DNA Fingerprinting / Q000191:economics* / Q000379:methods*; D004586:Electrophoresis; D006801:Humans; D016133:Polymerase Chain Reaction; D020080:Tandem Repeat Sequences
D004247:DNA