AgResearch Limited, Grasslands Research Centre, Palmerston North, New Zealand.
Methods Mol Biol. 2021;2369:27-40. doi: 10.1007/978-1-0716-1681-9_3.
We present a detailed method for extraction of high-molecular weight genomic DNA suitable for numerous DNA sequencing applications, and a straightforward in silico approach for reconstructing novel mitochondrial (mt) genomes directly from total genomic DNA extracts derived from next-generation sequencing (NGS) data sets. The in silico post-sequencing pipeline described is fast, accurate, and highly efficient, with modest memory requirements that can be performed using a standard desktop computer. The approach is particularly effective for obtaining mitochondrial genomes for species with little or no mitochondrial sequence information currently available and overcomes many of the limitations of traditional strategies. The described methodologies are also applicable for metagenomics sequencing from mixed or pooled samples containing multiple species and subsequent specific assembly of specific mitochondrial genomes.
我们提出了一种详细的方法，用于提取适合多种 DNA 测序应用的高分子量基因组 DNA，以及一种从下一代测序 (NGS) 数据集的总基因组 DNA 提取物中直接重建新型线粒体 (mt) 基因组的简单的计算机方法。所描述的计算机后测序管道快速、准确且高效，对内存的要求适中，可使用标准台式计算机执行。该方法特别适用于获取目前可用线粒体序列信息很少或没有的物种的线粒体基因组，并克服了传统策略的许多限制。所描述的方法也适用于来自含有多个物种的混合或汇集样品的宏基因组测序，以及随后对特定线粒体基因组的特定组装。
