Exploring the Impacts of Ordinary Laboratory Alterations During Forensic DNA Processing on Peak Height Variation,...

Impacts of validation design on DNA signal were explored, and the level of variation introduced by injection, capillary changes, amplification, and kit lot was surveyed by examining a set of replicate samples ranging in mass from 0.25 to 0.008 ng. The variations in peak height, heterozygous balance, dropout probabilities, and baseline noise were compared using common statistical techniques. Data indicate that amplification is the source of the majority of the variation observed in the peak heights, followed by capillary lots. The use of different amplification kit lots did not introduce variability into the peak heights, heterozygous balance, dropout, or baseline. Thus, if data from case samples run over a significant time period are not available during validation, the validation must be designed to, at a minimum, include the amplification of multiple samples of varying quantity, with known genotype, amplified and run over an extended period of time using multiple pipettes and capillaries.