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线粒体DNA控制区域从高度降解的骨遗骸中通过单复数下一代测序进行类型化

Diana C Vinueza-Espinosa¹, Daniel R Cuesta-Aguirre¹, Assumpció Malgosa¹

¹Research Group in Biological Anthropology, Biological Anthropology Unit, Department of Animal Biology, Vegetal Biology and Ecology, Universitat Autònoma de Barcelona, Bellaterra, Spain.

Electrophoresis

|June 28, 2023

概括

线粒体DNA控制区域 (mtDNA-CR) 的下一代测序 (NGS) 成功地从退化的骨遗骸中获取遗传数据. M3协议和内部管道为法医识别提供了更好的结果.

科学领域:

法医科学法医科学法医科学
遗传学是一个遗传学.
分子生物学分子生物学

背景情况:

退化的骨遗骸在法医调查中对核DNA分析提出了挑战.
线粒体DNA (mtDNA) 类型,特别是控制区域 (CR),为从受损样本中进行遗传鉴定提供了可行的替代方案.
与传统方法相比,下一代测序 (NGS) 技术简化了mtDNA-CR分析.

研究的目的:

评估PowerSeq CRM嵌套系统套件对高度降解的人类骨遗骸的mtDNA-CR类型的有效性.
为了比较不同的聚合酶链反应 (PCR) 协议,以优化mtDNA-CR恢复.
评估内部生物信息管道的性能与用于变种分析的商业软件相比.

主要方法:

使用了PowerSeq CRM嵌套系统套件与嵌套多重复合PCR用于mtDNA-CR放大和索引.
在41个来自不同时期的骨样本上测试了三个修改的PCR协议 (M1,M2,M3).
将内部生物信息管道与GeneMarker HTS软件进行比较,用于分析测序数据.

主要成果:

标准协议 (M1) 取得了有限的分析成功.
采用M3协议,包括35个PCR循环和延长的变性/延长时间,显著改善了从降解样本中恢复mtDNA-CR的情况.
混合基因配置和受损读取百分比作为污染指标,加强数据的解释,当一起分析.

结论:

使用PowerSeq CRM嵌套系统套件,M3协议有效地从高度降解的骨遗骸中获得mtDNA-CR配置文件.
公司内部的生物信息管道提供可靠的变体数据,与商业法医软件相比.
混合基和受损基的综合分析有助于识别潜在的污染,并改善法医遗传识别.

关键词:

没有NGS,没有NGS.PowerSeq CRM 嵌套系统套件嵌套系统套件我们的mtDNA mtDNA 协议优化协议优化骨样本骨样本骨样本

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