Volume 48, Issue 5 (September 2003)

    Development of an Alu-based, Real-Time PCR Method for Quantitation of Human DNA in Forensic Samples

    (Received 15 March 2003; accepted 15 March 2003)

    Published Online: September

    CODEN: JFSOAD

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    Abstract

    Determining the amount of human DNA extracted from a crime scene sample is an important step in DNA profiling. The forensic community relies almost entirely upon a technique (slot blot) to quantitate human DNA that is imprecise, time consuming, and labor intensive. We have previously described a method for quantitation of human DNA based on PCR amplification of a repetitive Alu sequence that uses a fluorescence plate reader. This manuscript describes and validates a variation of this assay using real-time PCR and SYBR® Green I for quantitation. The advantages of the real-time assay over the plate reader assay are: reduced hands-on time, lower assay cost, and a greater dynamic range. The main disadvantage is the cost of the real-time instrument. However, for those forensic laboratories with access to a real-time instrument, this Alu-based assay has a dynamic range of 16 ng to 1 pg, is sensitive, specific, fast, quantitative, and uses only 2 µL of sample.


    Stock #: JFS2002414

    ISSN: 0022-1198

    DOI: 10.1520/JFS2002414

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    Author
    Title Development of an Alu-based, Real-Time PCR Method for Quantitation of Human DNA in Forensic Samples
    Symposium , 0000-00-00
    Committee E30