Molecular assay for screening and quantifying dna in biological evidence: The modified Q-tat assay

Jon Wilson, Valerie Fuller, Gifty Benson, Denise Juroske, Eric Duvall, Jun Fu, Jane Pritchard, Robert W. Allen

Research output: Contribution to journalArticle

4 Scopus citations

Abstract

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.

Original languageEnglish
Pages (from-to)1050-1057
Number of pages8
JournalJournal of Forensic Sciences
Volume55
Issue number4
DOIs
StatePublished - 1 Jul 2010

Keywords

  • PCR inhibitors
  • STR analysis
  • amelogenin
  • capillary electrophoresis
  • forensic science
  • gender typing
  • human DNA quantitation

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