A 26plex Autosomal STR Assay to Aid Human Identity Testing*

Authors


  • *

    National Institute of Justice (NIJ) funded the work described here through interagency agreement 2003-IJ-R-029 to the NIST Office of Law Enforcement Standards.

  • Official Disclaimer: Contribution of the U.S. National Institute of Standards and Technology. Not subject to copyright. Certain commercial equipment, instruments, and materials are identified in order to specify experimental procedures as completely as possible. In no case does such identification imply a recommendation or endorsement by the National Institute of Standards and Technology nor does it imply that any of the materials, instruments, or equipment identified are necessarily the best available for the purpose. Points of view in this document are those of the authors and do not necessarily represent the official position or policies of the U.S. Department of Justice.

Additional information and reprint requests:
Carolyn R. Hill, M.S.
National Institute of Standards and Technology
Biochemical Science Division
100 Bureau Drive, Mail Stop 8311
Gaithersburg, MD 20899-8311
E-mail: becky.hill@nist.gov

Abstract

Abstract:  A short tandem repeat multiplex assay has been successfully developed with 25 autosomal loci plus the sex-typing locus amelogenin for a total of 26 amplified products in a single reaction. Primers for the loci were designed so that all of the amplicons present were distributed from 65 base pairs (bp) to less than 400 bp within a five-dye chemistry design with the fifth dye reserved for the sizing standard. A multiplex design strategy was developed to overcome challenges encountered in creating this assay. The limits of the multiplex were tested, resulting in the successful amplification of a wide range of genomic DNA sample concentrations from 2 ng to as low as 100 pg with 30 cycles of PCR. The 26plex has the potential to benefit the forensic community for reference sample testing and complex relationship evaluation.

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