Abstract
STR multiplexes remain the cornerstone of genotyping forensic samples. The PowerPlex® 16 HS System contains the core CODIS loci: D3S1358, D5S818, D7S820, D8S1179, D13S317,
D16S539, D18S51, D21S11, CSF1PO, FGA, TH01, TPOX, and vWA. Additional loci amplified
in the multiplex reaction are the sex-determinant locus, amelogenin, and two pentanucleotide
STR loci, Penta D and Penta E. The PowerPlex® 16 HS System is an updated version of the PowerPlex 16® System; while the primers and dyes remain unchanged, it introduces an enhanced buffer
system that includes hot-start Taq DNA polymerase and ensures robust performance. Due to the modification of the reaction
mix, a multi-laboratory developmental validation study was completed to document performance
capabilities and limitations for the revised assay. Data within this validation was
generated by eight laboratories and served as the basis for the following conclusions:
genotyping of single-source samples was consistent across a large range of template
DNA concentrations with most laboratories obtaining complete profiles at 62.5 pg. Mixture analyses showed that over 90% of minor alleles were detected at 1:9 ratios.
Optimum amplification cycle number was ultimately dependent on the sensitivity of
the detection instrument and could be adjusted to accommodate a range of DNA template
concentrations. Reaction conditions including volume and annealing temperature as
well as the concentrations of primers, Taq DNA polymerase, and magnesium were shown to be optimal and able to withstand moderate
variations without affecting multiplexed STR amplification. Finally, data from non-probative
samples and concordance studies showed consistent results when comparing the PowerPlex® 16 HS System with the PowerPlex® 16 System as well as other commercially available systems.
Keywords
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Article info
Publication history
Published online: November 09, 2009
Accepted:
October 2,
2009
Received in revised form:
September 15,
2009
Received:
August 4,
2009
Identification
Copyright
© 2009 Elsevier Ireland Ltd. Published by Elsevier Inc. All rights reserved.
