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Developmental Validation of Reduced-Size STR Miniplex Primer Sets*

Authors

  • Kerry L. Opel M.A.,

    1. International Forensic Research Institute, Florida International University, 11200 SW 8th Street, Miami, FL 33199.
    2. Ohio University Department of Chemistry and Biochemistry, 136 Clippinger Laboratories, Athens, Ohio 45701.
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  • Denise T. Chung Ph.D.,

    1. Ohio University Department of Chemistry and Biochemistry, 136 Clippinger Laboratories, Athens, Ohio 45701.
    2. Center for Neurological Diseases, Brigham & Women’s Hospital, Harvard Institutes of Medicine, 77 Avenue, Louis Pasteur Room, Boston, MA 02115-5817.
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  • Jiří Drábek Ph.D.,

    1. Ohio University Department of Chemistry and Biochemistry, 136 Clippinger Laboratories, Athens, Ohio 45701.
    2. Department of Biochemistry, Palacky University, Olomouc, CZ-783, Czech Republic.
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  • John M. Butler Ph.D.,

    1. Biochemical Science Division, National Institute of Standards and Technology, Gaithersburg, MD 20899–8311.
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  • Bruce R. McCord Ph.D.

    1. International Forensic Research Institute, Florida International University, 11200 SW 8th Street, Miami, FL 33199.
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  • *

    This project was supported under award no. 2005-MU-BX-K073 from the National Institute of Justice, and funded in part by the National Institute of Justice through interagency agreement 2003-IJ-R-029 with the NIST Office of Law Enforcement Standards.

Abstract

Abstract:  This paper describes a developmental validation study of three Miniplex sets covering 12 of the 13 CODIS loci. As these new sets will be used for the analysis of degraded and low level DNA, the validation studies were performed using 100–125 pg of DNA, the lowest input level at which peak balance, peak intensity, and allele consistency were stable. To demonstrate the applicability of the Miniplex sets to forensic casework, these validation studies were completed in accordance with the Scientific Working Group on DNA Analysis Methods (SWGDAM). A range of tests were performed including studies of concordance with standard multiplex kits, sensitivity and reproducibility, and PCR amplification conditions. Additionally, studies of mixtures, nonhuman and environmentally degraded DNA, and simulated forensic samples were performed. Our results demonstrate that Miniplex STR amplification procedures are a robust and sensitive tool for the analysis of degraded DNA.

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