Rasmussen David A, Noor Mohamed A F
Department of Biology, Duke University, Durham, NC 27708, USA.
BMC Genomics. 2009 Aug 18;10:382. doi: 10.1186/1471-2164-10-382.
The declining cost of DNA sequencing is making genome sequencing a feasible option for more organisms, including many of interest to ecologists and evolutionary biologists. While obtaining high-depth, completely assembled genome sequences for most non-model organisms remains challenging, low-coverage genome survey sequences (GSS) can provide a wealth of biologically useful information at low cost. Here, using a random pyrosequencing approach, we sequence the genome of the scuttle fly Megaselia scalaris and evaluate the utility of our low-coverage GSS approach.
Random pyrosequencing of the M. scalaris genome provided a depth of coverage (0.05x0.1x) much lower than typical GSS studies. We demonstrate that, even with extremely low-coverage sequencing, bioinformatics approaches can yield extensive information about functional and repetitive elements. We also use our GSS data to develop genomic resources such as a nearly complete mitochondrial genome sequence and microsatellite markers for M. scalaris.
We conclude that low-coverage genome surveys are effective at generating useful information about organisms currently lacking genomic sequence data.
DNA测序成本的不断下降使得基因组测序成为更多生物可行的选择,包括许多生态学家和进化生物学家感兴趣的生物。虽然为大多数非模式生物获得高深度、完全组装的基因组序列仍然具有挑战性,但低覆盖度的基因组调查序列(GSS)能够以低成本提供大量生物学上有用的信息。在此,我们采用随机焦磷酸测序方法对蚤蝇(Megaselia scalaris)的基因组进行测序,并评估我们的低覆盖度GSS方法的效用。
对蚤蝇基因组进行随机焦磷酸测序所提供的覆盖深度(0.05x - 0.1x)远低于典型的GSS研究。我们证明,即使是极低覆盖度的测序,生物信息学方法也能产生关于功能元件和重复元件的大量信息。我们还利用我们的GSS数据开发了基因组资源,如蚤蝇几乎完整的线粒体基因组序列和微卫星标记。
我们得出结论,低覆盖度基因组调查在为目前缺乏基因组序列数据的生物生成有用信息方面是有效的。