Journal Published Online: 01 September 2003
Volume 48, Issue 5

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

CODEN: JFSCAS

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.

Author Information

Nicklas, JA
VT Forensic Laboratory, Waterbury, VT
Buel, E
VT Forensic Laboratory, Waterbury, VT
Pages: 9
Price: $25.00
Related
Reprints and Permissions
Reprints and copyright permissions can be requested through the
Copyright Clearance Center
Details
Stock #: JFS2002414
ISSN: 0022-1198
DOI: 10.1520/JFS2002414