Development and validation of a rapid PCR method for the PowerPlex® 16 HS system for forensic DNA identification

• Published in: International journal of legal medicine Vol. 129; no. 4; pp. 715 - 723
• Main Authors: White, James, Hughes-Stamm, Sheree, Gangitano, David
• Format: Journal Article
• Language: English
• Published: Berlin/Heidelberg Springer Berlin Heidelberg 01.07.2015; Springer Nature B.V
• Subjects: DNA Fingerprinting - methods; Efficiency, Organizational; Female; Forensic Medicine; Forensic sciences; Genetic testing; Humans; Laboratories; Male; Medical Law; Medicine; Medicine & Public Health; Polymerase Chain Reaction - methods; Technical Note; Time Factors; Validation studies; United States > US; Short tandem repeats; PowerPlex; Forensic DNA; Rapid PCR
• ISSN: 0937-9827; 1437-1596
• DOI: 10.1007/s00414-014-1102-1

Currently, the amplification step of most forensic DNA profiling systems takes 3–4 h to complete. A decrease in the amplification time would allow for increased laboratory throughput, which may help reduce backlog when is due to limited cycling capacity. By using the SpeedSTAR™ HS Polymerase (Takara Bio, Otsu, Japan) in combination with the Veriti® (Applied Biosystems, Foster City, CA, USA) rapid thermal cycler, the amplification time for the PowerPlex® 16 HS (Promega, Madison, WI, USA) kit was reduced by 66 % (1 h). The sensitivity of this fast method was comparable to the standard system (0.13 ng). Although this rapid protocol showed an increase in average stutter ratios (2.8 %) and a decrease in average peak height ratios (7 %) across all loci when compared with standard conditions, it was able to consistently generate reliable DNA profiles. The results of this study indicate that the rapid protocol could be implemented in forensic laboratories with an optimal range of 0.25–2 ng of input DNA using appropriate analytical interpretation guidelines.