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Molecular Assay for Screening and Quantifying DNA in Biological Evidence: The Modified Q-TAT Assay

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A method is described for the quantitation of total human and male DNA. Q-TAT utilizes end-point, multiplex polymerase chain reaction (PCR) amplification of the amelogenin and SRY loci to quantify DNA and incorporates a cloned nonhuman template to detect PCR inhibition. Standard curves of fluorescence from amelogenin or SRY amplicons were generated from amplification of known amounts of NIST traceable SRM-female or SRM-male DNA. Curves showed good linearity up to 500 pg of SRM-template (R2 > 0.99) and reliably estimated total and male DNA content in casework samples. The nonhuman pRLnull template included in each PCR was a sensitive indicator of known PCR inhibitors including EDTA, hemin, blue denim dye, and humic acid. Finally, the SRY amplicon was a sensitive indicator of male DNA and, in mixtures, could reliably estimate male DNA present in an excess of female DNA. The Q-TAT multiplex is a reliable quantitation method for forensic DNA typing.
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Keywords: PCR inhibitors; STR analysis; amelogenin; capillary electrophoresis; forensic science; gender typing; human DNA quantitation

Document Type: Research Article

Affiliations: 1: Police Laboratory, Tulsa Police Department, Tulsa, OK. 2: Department of Forensic Science, Center for Health Sciences, Oklahoma State University, Tulsa, OK.

Publication date: 2010-07-01

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