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A 26plex Autosomal STR Assay to Aid Human Identity Testing

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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.

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

Document Type: Research Article

DOI: http://dx.doi.org/10.1111/j.1556-4029.2009.01110.x

Affiliations: Biochemical Science Division, National Institute of Standards and Technology, Gaithersburg, MD 20899-8311.

Publication date: September 1, 2009

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