Real-time polymerase chain reaction quantification of canine DNA.
The accurate quantification of target DNA is an important step in the short tandem repeat analysis of forensic biological samples. By utilizing quantification data to control the amount of template DNA in the polymerase chain reaction (PCR), forensic scientists can optimize testing and minimize the consumption of limited samples. The ability to identify and quantify target DNA in mixed-species samples is crucial when it may be overwhelmed by nontarget DNA, as in cases of dog attack. We evaluated two quantitative real-time PCR assays for dynamic range, species specificity, and inhibition by humic acid. While both assays proved to be highly sensitive and discriminating, the Melanocortin-1 Receptor (MC1R) gene Taqman assay had the advantages of a shorter run time, greater efficiency, and safer reagents. In its application to forensic casework, the MC1R assay has been advantageous for quantifying dog DNA in a variety of mixed-species samples and facilitating the successful profiling of individual dogs.
Evans
Jeffrey J
JJ
Veterinary Genetics Laboratory, Forensic Unit, School of Veterinary Medicine, University of California, Davis, CA 95616-8744, USA. jjevans@ucdavis.edu
Wictum
Elizabeth J
EJ
Penedo
M Cecilia T
MC
Kanthaswamy
Sreetharan
S
D000818:Animals; D002415:Cats; D002417:Cattle; D002614:Chelating Agents; D004247:DNA / Q000235:genetics*; D017931:DNA Primers; D015342:DNA Probes; D003670:Deer; D004285:Dogs / Q000235:genetics*; D005453:Fluorescence; D053803:Forensic Genetics / Q000379:methods; D006041:Goats; D006736:Horses; D006801:Humans; D006812:Humic Substances; D016133:Polymerase Chain Reaction / Q000379:methods*; D044102:Receptor, Melanocortin, Type 1 / Q000235:genetics*; D013045:Species Specificity
D002614:Chelating Agents; D017931:DNA Primers; D015342:DNA Probes; D006812:Humic Substances; D044102:Receptor, Melanocortin, Type 1; D004247:DNA