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학술연구/단체지원/교육 등 연구자 활동을 지속하도록 DBpia가 지원하고 있어요.
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연구자들이 자신의 연구와 전문성을 널리 알리고, 새로운 협력의 기회를 만들 수 있는 네트워킹 공간이에요.
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Recently, next generation sequencing (NGS) has received attention as the ultimategenotyping method to overcome the limitations of capillary electrophoresis (CE)-based short tandem repeat (STR) analysis, such as the limited number of STR locithat can be measured simultaneously using fluorescent-labeled primers and the maximumsize of STR amplicons. In this study, we analyzed 15 autosomal STR markersvia the NGS method and evaluated their effectiveness in STR analysis. Using maleand female standard DNA as single-sources and their 1:1 mixture, we sequentiallygenerated sample amplicons by the multiplex polymerase chain reaction (PCR)method, constructed DNA libraries by ligation of adapters with a multiplex identifier(MID), and sequenced DNA using the Roche GS Junior Platform. Sequencing datafor each sample were analyzed via alignment with pre-built reference sequences.
Most STR alleles could be determined by applying a coverage threshold of 20% forthe two single-sources and 10% for the 1:1 mixture. The structure of the STR in eachallele was accurately determined by examining the sequences of the target STRregion. The mixture ratio of the mixed sample was estimated by analyzing the coverageratios between assigned alleles at each locus and the reference/variant ratiosfrom the observed sequence variations. In conclusion, the experimental method usedin this study allowed the successful generation of NGS data. In addition, the NGS dataanalysis protocol enables accurate STR allele call and repeat structure determinationat each locus. Therefore, this approach using the NGS system will be helpful to interpretand analysis the STR profiles from singe-source and even mixed samples inforensic investigation.
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참고문헌 (21)
참고문헌 신청
Bamshad MJ / 2012 / Exome sequencing as a tool for Mendelian disease gene discovery / Nat Rev Genet 12 : 745 ~ 755
Berglund EC / 2011 / Next-generation sequencing technologies and applications for human genetic history and forensics / Investig Genet 2 : 23 ~
Bornman DM / 2012 / Short-read, high-throughput sequencing technology for STR genotyping / Biotechniques 0 : 1 ~ 6
Cho IS / 2012 / The human microbiome : at the interface of health and disease / Nat Rev Genet 13 : 260 ~ 270
Dalsgaard S / 2014 / Non-uniform phenotyping of D12S391 resolved by second generation sequencing / Forensic Sci Int Genet 8 : 195 ~ 199
Berglund EC / 2011 / Next-generation sequencing technologies and applications for human genetic history and forensics / Investig Genet 2 : 23 ~
Bornman DM / 2012 / Short-read, high-throughput sequencing technology for STR genotyping / Biotechniques 0 : 1 ~ 6
Cho IS / 2012 / The human microbiome : at the interface of health and disease / Nat Rev Genet 13 : 260 ~ 270
Dalsgaard S / 2014 / Non-uniform phenotyping of D12S391 resolved by second generation sequencing / Forensic Sci Int Genet 8 : 195 ~ 199
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