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

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

CODEN: JFSCAS

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.

Author Information

Leclair, B
Royal Canadian Mounted Police, Ottawa, Ontario, Canada Myriad Genetic Laboratories, Salt Lake City, UT
Sgueglia, JB
Suffolk County Crime Laboratory, Hauppauge, NY Massachusetts State Police Crime Laboratory, Sudbury, MA
Wojtowicz, PC
Minnesota BCA Forensic Science Laboratory, St. Paul, MN
Juston, AC
Suffolk County Crime Laboratory, Hauppauge, NY
Fregeau, CJ
Royal Canadian Mounted Police, Ottawa, Ontario, Canada
Fourney, RM
Royal Canadian Mounted Police, Ottawa, Ontario, Canada
Pages: 13
Price: $25.00
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Stock #: JFS2002182
ISSN: 0022-1198
DOI: 10.1520/JFS2002182