Part of the work was presented at the 59th Annual Meeting of the American Academy of Forensic Sciences, in San Antonio, TX, February 19–24, 2007, and at the 18th International Symposium on Human Identification, Hollywood, CA, October 2007. Some of the data is also presented in the Quantifiler® Duo DNA Quantification Kit User Manual.
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*
Article first published online: 27 JAN 2009
© 2009 Applied Biosystems Journal compilation © 2009 American Academy of Forensic Sciences
Journal of Forensic Sciences
Volume 54, Issue 2, pages 305–319, March 2009
How to Cite
Barbisin, M., Fang, R., O’Shea, C. E., Calandro, L. M., Furtado, M. R. and Shewale, J. G. (2009), 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. Journal of Forensic Sciences, 54: 305–319. doi: 10.1111/j.1556-4029.2008.00951.x
- Issue published online: 24 FEB 2009
- Article first published online: 27 JAN 2009
- Received 19 Jan. 2008; and in revised form 3 April 2008; accepted 20 April 2008.
- forensic science;
- DNA quantification;
- real-time PCR;
- DNA analysis;
- ribonuclease P RNA component H1;
- sex determining region Y;
- human DNA;
- human male DNA;
- DNA typing
Abstract: 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 (CT) 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.