title: A 26plex autosomal STR assay to aid human identity testing*.
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.
authors.affiliation: Biochemical Science Division, National Institute of Standards and Technology, Gaithersburg, MD 20899-8311, USA.
authors.forename: Carolyn R
authors.initials: CR
authors.lastname: Hill
authors.forename: John M
authors.initials: JM
authors.lastname: Butler
authors.forename: Peter M
authors.initials: PM
authors.lastname: Vallone
mesh_terms: D053523:Amelogenin / Q000235:genetics; D016172:DNA Fingerprinting / Q000379:methods*; D017931:DNA Primers; D018732:Forensic Anthropology; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D012984:Software; D020080:Tandem Repeat Sequences
chemical_list: D053523:Amelogenin; D017931:DNA Primers