ISSN: 0022-1198
CODEN: JFSCA
Published Online: 1 September 2003
Page Count: 9

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)

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.

Keywords:
forensic science, human DNA, DNA quantitation, Alu sequences, polymerase chain reaction, real time

Paper ID: JFS2002414
DOI: 10.1520/JFS2002414
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