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Developmental Validation of the Quantifiler ® Duo DNA Quantification Kit for Simultaneous Quantification of Total Human and Human Male DNA and Detection of PCR Inhibitors in Biological Samples *
Author(s) -
Barbisin Maura,
Fang Rixun,
O’Shea Cristin E.,
Calandro Lisa M.,
Furtado Manohar R.,
Shewale Jaiprakash G.
Publication year - 2009
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.1556-4029.2008.00951.x
Subject(s) - dna , dna profiling , computational biology , biology , chromatography , microbiology and biotechnology , chemistry , genetics
The Quantifiler ® Duo DNA Quantification kit enables simultaneous quantification of human DNA and human male DNA as well as detection of inhibitors of PCR in a single real‐time PCR well. Pooled human male genomic DNA is used to generate standard curves for both human (ribonuclease P RNA component H1) and human male (sex determining region Y) specific targets. A shift in the cycle threshold ( C T ) values for the internal positive control monitors the presence of PCR inhibitors in a sample. The assay is human specific and exhibits a high dynamic range from 0.023 to 50 ng/μL. In addition, the multiplex assay can detect as little as 25 pg/μL of human male DNA in the presence of a 1000‐fold excess of human female DNA. The multiplex assay provides assessment of the DNA extract and guidance for the selection of the appropriate AmpFℓSTR ® Amplification Kit to obtain interpretable short tandem repeat profiles.