Premium
Molecular Assay for Screening and Quantifying DNA in Biological Evidence: The Modified Q‐TAT Assay *
Author(s) -
Wilson Jon,
Fuller Valerie,
Benson Gifty,
Juroske Denise,
Duvall Eric,
Fu Jun,
Pritchard Jane,
Allen Robert W.
Publication year - 2010
Publication title -
journal of forensic sciences
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.715
H-Index - 96
eISSN - 1556-4029
pISSN - 0022-1198
DOI - 10.1111/j.1556-4029.2010.01371.x
Subject(s) - amelogenin , amplicon , microbiology and biotechnology , testis determining factor , dna , primer dimer , biology , multiplex , primer (cosmetics) , polymerase chain reaction , multiplex polymerase chain reaction , chemistry , genetics , gene , y chromosome , organic chemistry
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 ( R 2 > 0.99) and reliably estimated total and male DNA content in casework samples. The nonhuman pRL null 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.