Development of fish diversity studies by next-generation mitogenome sequencing: From Sanger method to massively parallel method

全线粒体基因组（有丝分裂基因组）序列对于解决包括鱼类在内的动物的系统发育非常有用。本研究旨在通过“下一代”大规模平行测序技术，建立有效的程序，从许多样品中获得有丝分裂基因组数据，包括降解的样品。对于条件良好的样品，long-PCR扩增/纯化和平行标记测序同时成功地提供了许多样品的有丝分裂基因组数据，这表明这是一种相对容易的方法。对于降解样品，使用双标引法捕获有丝分裂基因组的许多片段。因此，我们得出的结论是，通过使用下一代测序技术，已经建立了有效的程序，可以从许多样品（包括降解样品）中获得大量有丝分裂基因组数据。

Whole mitochondrial genome (mitogenome) sequences are extremely useful for resolving phylogenies of animals including fishes. This study aims to establish efficient procedures to obtain mitogenome data from many samples including degraded ones by using "next-generation" massively parallel sequencing technologies. For samples in good condition, long-PCR amplification/purification and parallel tagged sequencing successfully provided mitogenome data from many samples simultaneously, suggesting that this is a rather easier method for such samples. For degraded samples, many segments from mitogenome were successfully captured by using the capture-on-beads method with double indexing. Thus, we conclude that efficient procedures have been established to have large mitogenome data from many samples including degraded ones by using next-generation sequencing technologies.