STR DNA Typing: Increased Sensitivity and Efficient Sample Consumption Using Reduced PCR Reaction Volumes

    Volume 48, Issue 5 (September 2003)

    ISSN: 0022-1198

    CODEN: JFSOAD

    Published Online: 1 September 2003

    Page Count: 13


    (Received 4 April 2003; accepted 30 March 2003)

    Abstract

    Improvements in detection limits/sensitivity and lower sample consumption are potential benefits of reducing PCR reaction volumes used in forensic DNA typing of crime scene samples. This premise was studied first with experimental mixtures and a nine-loci megaplex, which demonstrated stochiometric amplification and accurate detection. Next, adjudicated casework samples were subjected to amplification under 15 different template DNA to PCR reaction volume ratios. Reduction of PCR reaction volume and DNA down to 10 µL and 0.500 ng, respectively, produced identical profiles with the same signal intensity and heterozygous allele peak height ratio (HR). Reduction to 5 µL and 0.063 ng yielded HR values that were slightly affected in one to three STR loci. PCR reaction volume reduction can enhance detection and sensitivity while reducing the consumption of irreplaceable crime scene samples.


    Paper ID: JFS2002182

    DOI: 10.1520/JFS2002182

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    Title STR DNA Typing: Increased Sensitivity and Efficient Sample Consumption Using Reduced PCR Reaction Volumes
    Symposium , 0000-00-00
    Committee E30