A 26plex autosomal STR assay to aid human identity testing*.
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.
Hill
Carolyn R
CR
Biochemical Science Division, National Institute of Standards and Technology, Gaithersburg, MD 20899-8311, USA.
Butler
John M
JM
Vallone
Peter M
PM
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
D053523:Amelogenin; D017931:DNA Primers