Schwartz Katja, Sherlock Gavin
Department of Genetics, Stanford University Medical School, Stanford, California 94305-5120.
Cold Spring Harb Protoc. 2016 Oct 3;2016(10):2016/10/pdb.top077651. doi: 10.1101/pdb.top077651.
The original yeast genome sequencing project was a monumental task, spanning several years, which resulted in the first sequenced eukaryotic genome. The 12 Mbp reference sequence was generated from yeast strain S288c and was of extremely high quality. In the years since it was published, sequencing technology has advanced apace, such that it is within the reach of most labs to sequence yeast strains of interest almost as a matter of standard practice, either via core facilities at their institution or through commercial sequencing services. Because of the availability of the high-quality reference sequence (which itself has received approximately 1500 updates derived from high-throughput sequencing data), reliable identification of differences between a strain of interest and the reference is relatively straightforward, at least for the nonrepetitive regions of the genome. In this introduction, we describe current high-throughput sequencing technology and methods for analysis of the resulting data.
最初的酵母基因组测序项目是一项耗时数年的艰巨任务,其成果是首个被测序的真核生物基因组。这条12兆碱基对的参考序列由酵母菌株S288c生成,质量极高。自其发表后的数年里,测序技术飞速发展,以至于现在大多数实验室几乎都能将对感兴趣的酵母菌株进行测序作为一种标准操作来完成,要么通过所在机构的核心设施,要么借助商业测序服务。由于高质量参考序列的存在(其本身已根据高通量测序数据进行了约1500次更新),至少对于基因组的非重复区域而言,可靠地识别感兴趣菌株与参考菌株之间的差异相对简单。在本引言中,我们描述了当前的高通量测序技术以及分析所得数据的方法。