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Updated Validation of Promega's PowerPlex 16 System: High Throughput Databasing Under Reduced PCR Volume Conditions on Applied Biosystem's 96 Capillary 3730xl DNA Analyzer

NCJ Number
225598
Journal
Journal of Forensic Sciences Volume: 53 Issue: 6 Dated: November 2008 Pages: 1353-1357
Author(s)
Rita Spathis B.S.; J. Koji Lum Ph.D.
Date Published
November 2008
Length
5 pages
Annotation
This report presents an updated validation of the PowerPlex 16 System from Promega Corporation on Applied Biosystem’s 96 capillary 3730xl DNA Analyzer.
Abstract
The validation concluded that the 3730xl PowerPlex 16 System is robust and accurate as well as optimally suited to DNA databasing and addressing the backlog of DNA samples facing many State laboratories. The reduction in reaction volume combined with the greater dynamic range of the 3730xl platform compared to previous generation DNA analyzers resulted in an overall increase in signal and assay sensitivity. In addition, the capacity of this extended validation to yield accurate profiles over a wide range of input template means fewer reruns due to too little or too much DNA template, further increasing the throughput capability of the assay. These modifications did not adversely affect heterozygous locus peak ratios, which remained above 60 percent. The quality of the data is reflected in the ability to reproducibly obtain complete and accurate profiles using nonprobative samples with a 99.8 percent (463/464 loci correctly identified) success rate. Coupled with the DNA concentration titration experiment, these data suggest that a wide range of field-collected samples would also result in accurate profiles. Male 9948 and female 9947A DNA standards were obtained from Promega Corporation, Madison, WI, as components of the PowerPlex Y (Lot # 216559) and PowerPlex 16 (Lot #s 218993, 222284) kits, respectively. In addition to describing the DNA samples, the description of materials and methods addresses PCR components; PCR amplification and thermal cycling parameters; and sample electrophoresis, bin settings, and data analysts. 5 figures and 14 references

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