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Developmental validation of the Quantifiler® HP and Trio Kits for human DNA quantification in forensic samples

Published:December 28, 2015DOI:https://doi.org/10.1016/j.fsigen.2015.12.007

      Highlights

      • The Quantifiler® HP and Quantifiler® Trio DNA Quantification Kits were designed to quantify human genomic DNA in forensic casework samples.
      • The kits are primarily intended as a preliminary screening and quantification step to performing analyses such as STR (short tandem repeat) genotyping.
      • The kits were designed to improve upon the sensitivity, robustness and time-to-result of earlier-generation DNA quantification kits, to be more aligned with current-generation STR kits.
      • This paper presents the results of developmental validation studies of the Quantifiler HP and Quantifiler Trio Kits.

      Abstract

      The quantification of human genomic DNA is a necessary first step in the DNA casework sample analysis workflow. DNA quantification determines optimal sample input amounts for subsequent STR (short tandem repeat) genotyping procedures, as well as being a useful screening tool to identify samples most likely to provide probative genotypic evidence. To better mesh with the capabilities of newest-generation STR analysis assays, the Quantifiler® HP and Quantifiler® Trio DNA Quantification Kits were designed for greater detection sensitivity and more robust performance with samples that contain PCR inhibitors or degraded DNA. The new DNA quantification kits use multiplex TaqMan® assay-based fluorescent probe technology to simultaneously quantify up to three human genomic targets, allowing samples to be assessed for total human DNA, male contributor (i.e., Y-chromosome) DNA, as well as a determination of DNA degradation state. The Quantifiler HP and Trio Kits use multiple-copy loci to allow for significantly improved sensitivity compared to earlier-generation kits that employ single-copy target loci. The kits’ improved performance provides better predictive ability for results with downstream, newest-generation STR assays, and their shortened time-to-result allows more efficient integration into the forensic casework analysis workflow.

      Keywords

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