Developmental Validation of Feline, Bovine, Equine, and Cervid Quantitative PCR Assays

Authors

  • Christina D. Lindquist M.Sc.,

    1. Veterinary Genetics Laboratory, Forensic Unit, School of Veterinary Medicine, University of California, One Shields Avenue, Davis, CA 95616.
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  • Jeffery J. Evans B.Sc.,

    1. Genetics Graduate Group, University of California, One Shields Avenue, Davis, CA 95616.
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  • Elizabeth J. Wictum B.Sc.

    1. Veterinary Genetics Laboratory, Forensic Unit, School of Veterinary Medicine, University of California, One Shields Avenue, Davis, CA 95616.
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  • Presented at the 60th Annual Meeting of the American Academy of Forensic Sciences, February 18–23, 2008, in Washington, DC. Also presented in part at the 18th International Symposium on Human Identification, October 2–3, 2007, in Hollywood, CA.

Additional information—reprints not available from author:
Christina D. Lindquist, M.S.F.S.
Veterinary Genetics Laboratory
Forensic Unit
University of California at Davis
One Shields Avenue
Davis, CA 95616-8744
E-mail: cdlindquist@ucdavis.edu

Abstract

Abstract:  Accurate DNA quantification is essential for optimizing DNA testing and minimizing sample consumption. Real-time quantitative polymerase chain reaction (qPCR) assays have been published for human and canine nuclear DNA, and the need for quantifying other forensically important species was evident. Following the strategy employed for the canine qPCR assay, we developed individual assays to accurately quantify feline, bovine, equine, and cervid nuclear DNA. Each TaqMan-based assay incorporates a genus-specific probe targeting the Melanocortin-1 Receptor gene and includes a piece of synthetic DNA that acts as an internal PCR control for detecting inhibition. Developmental validations were carried out following the revised guidelines of the Scientific Working Group on DNA Analysis Methods with modifications necessary for validation of nonhuman qPCR assays. All assays demonstrated the specificity, sensitivity, stability, reproducibility, accuracy, and precision required for forensic casework. The application of these assays to animal forensic DNA analysis has both conserved laboratory resources and improved genotyping results.

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