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*



This article is corrected by:

  1. Errata: Errata Volume 54, Issue 6, 1502, Article first published online: 23 October 2009
  2. Errata: Erratum Volume 57, Issue 1, 282, Article first published online: 4 January 2012

  • *

    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.

Additional information and reprint requests:
Jaiprakash G. Shewale, Ph.D.
Applied Biosystems
850 Lincoln Centre Drive, Mail Stop 402
Foster City, CA 94404


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.