TY - JOUR
T1 - An evaluation of the PowerSeq™ Auto System
T2 - A multiplex short tandem repeat marker kit compatible with massively parallel sequencing
AU - Zeng, Xiangpei
AU - King, Jonathan
AU - Hermanson, Spencer
AU - Patel, Jaynish
AU - Storts, Douglas R.
AU - Budowle, Bruce
PY - 2015/8/3
Y1 - 2015/8/3
N2 - 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.
AB - 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.
KW - Massively parallel sequencing (MPS)
KW - MiSeq
KW - Mixture
KW - PowerSeq Auto System
KW - Short tandem repeat (STR)
KW - Single nucleotide polymorphism (SNP)
UR - http://www.scopus.com/inward/record.url?scp=84938375264&partnerID=8YFLogxK
U2 - 10.1016/j.fsigen.2015.07.015
DO - 10.1016/j.fsigen.2015.07.015
M3 - Article
C2 - 26240968
AN - SCOPUS:84938375264
SN - 1872-4973
VL - 19
SP - 172
EP - 179
JO - Forensic Science International: Genetics
JF - Forensic Science International: Genetics
M1 - 1393
ER -