单环刺螠线粒体基因组全序列的获得——长PCR 结合鸟枪法测序

由于具有单基因所不可比拟的优势,线粒体基因组现已成为后生动物种群遗传和分子系统发育研究中一个重要的信息来源。本研究选取螠虫动物的代表物种——单环刺螠(Urechis unicinctus),详细阐述了利用长PCR方法扩增其线粒体DNA,获得约15 kb的扩增产物,进而构建shotgun文库,最终成功获得单环刺螠线粒体基因组全序列的流程。本实验流程样品需求量少、设备要求低、简便快捷。

Complete mitochondrial genome sequence of Urechis unicinctus accomplished by long PCR combined with shotgun sequencing

Due to their many advantages over single genes, mitochondrial genomes have become an important source of information in metazoan population genetics and molecular phylogenetic studies. In this study, Urechis unicinctus, a represent species of Echiura, was chosen to be as an example to clearly elaborate the process of long PCR amplification of its mitochondrial DNA. The long PCR amplified product was about 15kb. Then the shotgun library was constructed and the entire mitochondrial genome sequence was successfully obtained. The results show that the long PCR-based method is less demand of devices and amounts of tissue compared with physical separation method.