title: An evaluation of the PowerSeq™ Auto System: A multiplex short tandem repeat marker kit compatible with massively parallel sequencing.
abstract: Capillary electrophoresis (CE) and multiplex amplification with fluorescent tagging have been routinely used for STR typing in forensic genetics. However, CE-based methods restrict the number of markers that can be multiplexed simultaneously and cannot detect any intra-repeat variations within STRs. Several studies already have indicated that massively parallel sequencing (MPS) may be another potential technology for STR typing. In this study, the prototype PowerSeq(™) Auto System (Promega) containing the 23 STR loci and amelogenin was evaluated using Illumina MiSeq. Results showed that single source complete profiles could be obtained using as little as 62 pg of input DNA. The reproducibility study showed that the profiles generated were consistent among multiple typing experiments for a given individual. The mixture study indicated that partial STR profiles of the minor contributor could be detected up to 19:1 mixture. The mock forensic casework study showed that full or partial profiles could be obtained from different types of single source and mixture samples. These studies indicate that the PowerSeq Auto System and the Illumina MiSeq can generate concordant results with current CE-based methods. In addition, MPS-based systems can facilitate mixture deconvolution with the detection of intra-repeat variations within length-based STR alleles.
authors.affiliation: Institute of Applied Genetics, Department of Molecular and Medical Genetics, University of North Texas Health Science Center, 3500 Camp Bowie Blvd., Fort Worth, Texas 76107, USA. Electronic address: Xiangpei.Zeng@live.unthsc.edu.
authors.forename: Xiangpei
authors.initials: X
authors.lastname: Zeng
authors.affiliation: Institute of Applied Genetics, Department of Molecular and Medical Genetics, University of North Texas Health Science Center, 3500 Camp Bowie Blvd., Fort Worth, Texas 76107, USA.
authors.forename: Jonathan
authors.initials: J
authors.lastname: King
authors.affiliation: Promega Corporation, 2800 Woods Hollow Road, Madison, WI 53711, USA.
authors.forename: Spencer
authors.initials: S
authors.lastname: Hermanson
authors.affiliation: Promega Corporation, 2800 Woods Hollow Road, Madison, WI 53711, USA.
authors.forename: Jaynish
authors.initials: J
authors.lastname: Patel
authors.affiliation: Promega Corporation, 2800 Woods Hollow Road, Madison, WI 53711, USA.
authors.forename: Douglas R
authors.initials: DR
authors.lastname: Storts
authors.affiliation: Institute of Applied Genetics, Department of Molecular and Medical Genetics, University of North Texas Health Science Center, 3500 Camp Bowie Blvd., Fort Worth, Texas 76107, USA; Center of Excellence in Genomic Medicine Research (CEGMR), King Abdulaziz University, Jeddah, Saudi Arabia.
authors.forename: Bruce
authors.initials: B
authors.lastname: Budowle
mesh_terms: D005260:Female; D053803:Forensic Genetics; D005819:Genetic Markers; D059014:High-Throughput Nucleotide Sequencing / Q000295:instrumentation*; D006801:Humans; D008297:Male; D018895:Microsatellite Repeats / Q000235:genetics*; D016133:Polymerase Chain Reaction; D015203:Reproducibility of Results
chemical_list: D005819:Genetic Markers
keywords: Massively parallel sequencing (MPS); MiSeq; Mixture; PowerSeq Auto System; Short tandem repeat (STR); Single nucleotide polymorphism (SNP)