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首页> 外文期刊>Electrophoresis: The Official Journal of the International Electrophoresis Society >Multiplex pyrosequencing of InDel markers for forensic DNA analysis
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Multiplex pyrosequencing of InDel markers for forensic DNA analysis

机译：用于法医DNA分析的吲哚标记的多重发酵杆菌

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摘要

The capillary electrophoresis (CE) technology is commonly used for fragment length separation of markers in forensic DNA analysis. In this study, pyrosequencing technology was used as an alternative and rapid tool for the analysis of biallelic InDel (insertion/deletion) markers for individual identification. The DNA typing is based on a subset of the InDel markers that are included in the Investigator ? DIPplex Kit, which are sequenced in a multiplex pyrosequencing analysis. To facilitate the analysis of degraded DNA, the polymerase chain reaction (PCR) fragments were kept short in the primer design. Samples from individuals of Swedish origin were genotyped using the pyrosequencing strategy and analysis of the Investigator ? DIPplex markers with CE. A comparison between the pyrosequencing and CE data revealed concordant results demonstrating a robust and correct genotyping by pyrosequencing. Using optimal marker combination and a directed dispensation strategy, five markers could be multiplexed and analyzed simultaneously. In this proof‐of‐principle study, we demonstrate that multiplex InDel pyrosequencing analysis is possible. However, further studies on degraded samples, lower DNA quantities, and mixtures will be required to fully optimize InDel analysis by pyrosequencing for forensic applications. Overall, although CE analysis is implemented in most forensic laboratories, multiplex InDel pyrosequencing offers a cost‐effective alternative for some applications.
机译：毛细管电泳（CE）技术通常用于法医DNA分析中标志物的片段长度分离。在本研究中，焦塞节目技术被用作分析双射击靛蓝（插入/删除）标记的替代和快速工具，以进行个别识别。 DNA键入基于调查员中包含的indel标记的子集？ DIPPLEX试剂盒，其在多重焦点测序分析中测序。为了便于降解DNA的分析，在引物设计中保持聚合酶链反应（PCR）片段。使用焦肌策略和调查员的分析，瑞典源自源性的样品进行基因分型？与CE的冻倒像标记。焦点测序和CE数据之间的比较显示了通过焦肌肉进行稳健和正确的基因分型的一致性结果。使用最佳标记组合和定向分配策略，可以同时复用五个标记。在这种原则上的研究中，我们证明了多重诱导焦素测序分析是可能的。然而，需要进一步研究降解的样品，较低的DNA量和混合物，以通过对法医应用进行焦肌肉进行全面优化诱导分析。总的来说，尽管在大多数法医实验室实施了CE分析，但多重indel焦点测序为某些应用提供了一种经济高效的替代方案。

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