• forensic science;
  • short tandem repeat;
  • DNA;
  • miniSTR;
  • multiplex;
  • polymerase chain reaction;
  • D1GATA113;
  • D1S1627;
  • D1S1677;
  • D2S441;
  • D2S1776;
  • D3S3053;
  • D3S4529;
  • D4S2364;
  • D4S2408;
  • D5S2500;
  • D6S474;
  • D6S1017;
  • D8S1115;
  • D9S1122;
  • D9S2157;
  • D10S1248;
  • D10S1435;
  • D11S4463;
  • D12ATA63;
  • D14S1434;
  • D17S974;
  • D17S1301;
  • D18S853;
  • D20S482;
  • D20S1082;
  • D22S1045

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.