《生命科学》 2016, 28(1): 112-117
摘 要:摘 要:综述了高通量测序技术在线粒体全基因组测序中的策略,利用该技术对线粒体全基因组进行序列测定的方法可以归纳为两种,一种是先对目标mtDNA 进行富集,包括mtDNA 的提取纯化,目标区域PCR扩增法以及特异性探针杂交富集法( 可分为基于微阵列和基于PCR 探针的杂交富集法),然后对富集出的线粒体DNA 进行高通量测序;另一种是先从待测样本的基因组高通量数据中挖掘出线粒体基因组序列信息,之后利用诱饵序列或者近缘物种的线粒体全基因组参考序列,使用软件MITObim 对其进行组装。此外,还给出了线粒体高通量测序的优化流程图和介绍了混合样品的线粒体高通量测序策略。
Abstract: Abstract: This paper reviews the application of high-throughput sequencing technologies in the mitochondrial genomes. Two methods using high-throughput sequencing technology are included. The first method needs to enrich mtDNA from the sample, including extraction and purification of mtDNA, PCR amplicon-based, specific-probe hybridization (microarray-based and PCR-generation probe), then the enriched mtDNA was sequenced. For the second method, mitochondrial genome sequences were extracted from genome at first, then they were assembled using bait sequence or the mitochondrial genome sequence of related species as a reference, with software MITObim. In addition, the optimization flow chart of mitogenome sequencing using high-throughput sequencing technology was given in this paper. Finally, this paper also introduces the sequencing strategy of mixed sample.