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法医学杂志 2018, Vol. 34 Issue (5) :520-525    DOI: 10.12116/j.issn.1004-5619.2018.05.017
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基于Ion Torrent PGMTM测序平台的CSF1PO和D18S51基因座分析
杨仪尊1,2,平  原2
1. 复旦大学生命科学学院,上海 200438; 2. 上海市公安局物证鉴定中心 法医物证学现场应用技术公安部重点实验室 上海市现场物证重点实验室,上海 200083
Analysis of CSF1PO and D18S51 Loci Based on Ion Torrent PGMTM Platform
YANG Yi-zun1,2, PING Yuan2
1. School of Life Sciences, Fudan University, Shanghai 200438, China; 2. Shanghai Key Laboratory of Crime Scene Evidence, Key Laboratory of Forensic Evidence and Science Technology, Ministry of Public Security, Institute of Forensic Science, Shanghai Public Security Bureau, Shanghai 200083, China

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摘要 目的 应用二代测序(next generation sequencing,NGS)技术对CSF1PO和D18S51基因座进行分析检测,研究CSF1PO和D18S51基因座的序列多态性。 方法 采集165例中国汉族无关个体外周血样,应用QIAamp DNA Mini试剂盒提取样本DNA,Ion Plus Fragment Library试剂盒构建文库,在Ion Torrent PGMTM测序平台上进行DNA序列测定,针对新发现的等位基因进行Sanger测序验证。应用Torrent SuiteTM v5.0.2和Integrative Genomics Viewer软件分析数据,进行基因型鉴定和频率统计,运用PowerState v12软件对数据进行统计学处理。 结果 应用NGS技术同时获得了CSF1PO和D18S51基因座长度多态性和序列多态性,在CSF1PO基因座中,发现了1个新的基因型,在D18S51基因座中,发现2个新的基因型。采用Sanger测序法对NGS技术检测新发现的等位基因进行验证,验证结果一致。 结论 应用二代测序技术可检测CSF1PO和D18S51基因座核心重复序列的结构,提高基因座的识别效能。
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杨仪尊
平原
关键词法医遗传学   序列分析   DNA   CSF1PO   D18S51   二代测序   Ion Torrent PGMTM测序系统     
Abstract: Objective To analyse and detect CSF1PO and D18S51 loci by next generation sequencing (NGS) technology for the study on their sequence polymorphism. Methods The peripheral blood samples were collected from 165 unrelated individuals of Chinese Han population. DNA samples were obtained by QIAamp DNA Mini kit. The library was constructed by Ion Plus Fragment Library. DNA sequencing analysis was performed on Ion Torrent PGMTM Platform. The newfound alleles were verified by Sanger sequencing. Data were analysed by Torrent SuiteTM v5.0.2 and Integrative Genomics Viewer for the genotype identification and frequency count. The data were analysed statistically by PowerState v12. Results The length and sequence polymorphisms of CSF1PO and D18S51 loci were simultaneously obtained by NGS technology. A new genotype was found on CSF1PO locus, and two new genotypes on D18S51 locus. Sanger sequencing was used to verify the newfound alleles found by NGS technology, and the results of verification showed consistency. Conclusion The structure of core repeats on CSF1PO and D18S51 loci was detected by NGS in this study for the improvement of the identifying performance of locus.
Keywordsforensic genetics   sequence analysis, DNA   CSF1PO   D18S51   next generation sequencing   Ion Torrent PGMTM sequencing system     
About author: 杨仪尊(1988—),女,硕士研究生,主要从事法医物证学检验;E-mail:shnuyyz@163.com
引用本文:   
杨仪尊,平原.基于Ion Torrent PGMTM测序平台的CSF1PO和D18S51基因座分析[J]  法医学杂志, 2018,34(5): 520-525
YANG Yi-zun, PING Yuan.Analysis of CSF1PO and D18S51 Loci Based on Ion Torrent PGMTM Platform[J]  法医学杂志, 2018,34(5): 520-525
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