Developmental Validation of a Real-Time Quantitative PCR Assay for Automated Quantification of Human DNA

    Volume 48, Issue 5 (September 2003)

    ISSN: 0022-1198

    CODEN: JFSOAD

    Published Online: 1 September 2003

    Page Count: 6


    (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.


    Paper ID: JFS2002440

    DOI: 10.1520/JFS2002440

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    Title Developmental Validation of a Real-Time Quantitative PCR Assay for Automated Quantification of Human DNA
    Symposium , 0000-00-00
    Committee E30