Volume 42, Issue 1 (January 1997)

    Co-amplification of the Amelogenin and HLA DQα Genes: Optimization and Validation

    (Received 22 March 1996; accepted 27 May 1996)

    CODEN: JFSOAD

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    Abstract

    An optimized PCR-based system allowing the coamplification of the HLA DQα and the amelogenin genes has been developed and validated, enabling simultaneous identity testing and sex determination. Sensitivities below 100 pg of human DNA were obtained, using a convenient, high resolution agarose gel system and ethidium bromide staining. Comparison of several co-amplification methods revealed that the best sensitivities and most consistent results were obtained using a hotstart technique employing an inactivating antibody to Taq polymerase. HLA DQα typing results were reliably obtained using the co-amplification process. The sensitivity and ease of this system rendered it directly applicable to forensic analyses. The optimized techniques described here have been validated and successfully applied to forensic cases including People vs. Trujillo, in which the California Superior Court accepted these techniques as scientifically reliable and admissible. Work currently in progress has demonstrated that the described protocol may also be used to co-amplify the amelogenin gene with the AmpliType® PM (Polymarker) system, allowing identity testing at six loci in addition to sex determination.


    Author Information:

    Challed-Spong, A
    Director and Criminalist, Technical Associates, Inc., Altadena, CA

    Johnson, EA
    Director of DNA Laboratory, Harris County Medical Examiner's Office, Houston, TX

    Taylor, MS
    Director and Criminalist, Technical Associates, Inc., Altadena, CA


    Stock #: JFS14082J

    ISSN: 0022-1198

    DOI: 10.1520/JFS14082J

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    Title Co-amplification of the Amelogenin and HLA DQα Genes: Optimization and Validation
    Symposium , 0000-00-00
    Committee E30