McKusick-Nathans Institute of Genetic Medicine, Johns Hopkins School of Medicine, Baltimore, Maryland, USA.
PLoS Comput Biol. 2012;8(10):e1002737. doi: 10.1371/journal.pcbi.1002737. Epub 2012 Oct 25.
We describe methods for rapid sequencing of the entire human mitochondrial genome (mtgenome), which involve long-range PCR for specific amplification of the mtgenome, pyrosequencing, quantitative mapping of sequence reads to identify sequence variants and heteroplasmy, as well as de novo sequence assembly. These methods have been used to study 40 publicly available HapMap samples of European (CEU) and African (YRI) ancestry to demonstrate a sequencing error rate <5.63×10(-4), nucleotide diversity of 1.6×10(-3) for CEU and 3.7×10(-3) for YRI, patterns of sequence variation consistent with earlier studies, but a higher rate of heteroplasmy varying between 10% and 50%. These results demonstrate that next-generation sequencing technologies allow interrogation of the mitochondrial genome in greater depth than previously possible which may be of value in biology and medicine.
我们描述了快速测序整个人类线粒体基因组（mtgenome）的方法，这些方法包括长距离 PCR 对 mtgenome 的特异性扩增、焦磷酸测序、对序列读数进行定量映射以识别序列变体和异质性，以及从头序列组装。这些方法已被用于研究 40 个公开的欧洲（CEU）和非洲（YRI）祖先的 HapMap 样本，以证明测序错误率 <5.63×10(-4)，CEU 的核苷酸多样性为 1.6×10(-3)，YRI 的核苷酸多样性为 3.7×10(-3)，与早期研究一致的序列变异模式，但异质性的发生率在 10%到 50%之间更高。这些结果表明，下一代测序技术允许比以前更深入地研究线粒体基因组，这在生物学和医学中可能具有价值。
