ISSN: 0022-1198
CODEN: JFSCA
Published Online: 1 September 2003
Page Count: 6

Developmental Validation of a Real-Time Quantitative PCR Assay for Automated Quantification of Human DNA
(Received 19 April 2003; accepted 5 April 2003)

Abstract

Our laboratory has developed an automated real-time quantitative PCR assay for detecting human DNA. The assay utilizes an inhouse, custom-designed TaqMan®-MGB sequence-specific probe (CFS-rHumRT) and the ABD 7900HT SDS platform. Developmental validation has followed TWGDAM (1) guidelines and demonstrates that the assay is primate specific, is highly sensitive, yields consistent results, and works with human DNA extracted from a variety of body fluid stains. When operating within the dynamic range of the system using high-quality DNA samples, the technique yields similar quantification results to our current QuantiBlot™ assay with the added benefit of time saving through automation. Furthermore, the QPCR assay identifies how much amplifiable DNA is in a sample and thus has the potential to predict PCR success in downstream applications such as STR analysis.

Keywords:
forensic science, genomic DNA, quantification, real-time PCR, ABI 7900 Sequence Detection System, automation

Paper ID: JFS2002440
DOI: 10.1520/JFS2002440
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