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


  • *

    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.

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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:  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.