Since 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, hoping to reduce 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 percent, peak height imbalances improved, and drop-out was eliminated in 75.8 percent of samples. Only eight of 467 case samples required re-amplification, a success rate of 94 percent was observed, and the repeat rate decreased significantly. Finally, a DNA input of 0.25–1.0 ng is ideal for half-reaction amplification. (publisher abstract modified)
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