Whole-Genome Sequencing of Yeast Cells.
Whole-Genome Sequencing of Yeast Cells.
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DOI：
10.1002/cpmb.103
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发表时间：
2019-09-01
影响因子：
--
通讯作者：
Winston, Fred
中科院分区：
文献类型：
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作者：
Gopalakrishnan, Rajaraman;Winston, Fred
芽殖酵母（酿酒酵母）已广泛用于基本细胞功能的遗传研究。酵母突变体的分离和分析是一种常用的和强大的技术，以确定参与感兴趣的过程的基因。此外，野生酵母菌株之间的自然遗传变异进行了研究，通过数量性状基因座作图分析多基因性状。全基因组测序，通常与批量分离分析相结合，是一种强大的技术，有助于确定导致表型的突变的身份。在这里，我们描述了用于构建S的库的协议。酿酒酵母全基因组测序。我们还提出了一个生物信息学管道，使用全基因组测序数据来确定酵母菌株中的遗传变异。该流水线也可用于粟酒裂殖酵母突变体的分析。John Wiley & Sons，Inc._关于我们基本方案1：基本方案3：剪切基因组DNA用于文库制备基本方案4：DNA文库的构建和扩增支持方案1：退火寡核苷酸用于形成Y-接头支持方案2：使用SPRI珠粒进行大小选择和净化基本方案5：从测序数据鉴定基因组变体。
The budding yeast, Saccharomyces cerevisiae, has been widely used for genetic studies of fundamental cellular functions. The isolation and analysis of yeast mutants is a commonly used and powerful technique to identify the genes that are involved in a process of interest. Furthermore, natural genetic variation among wild yeast strains has been studied for analysis of polygenic traits by quantitative trait loci mapping. Whole-genome sequencing, often combined with bulk segregant analysis, is a powerful technique that helps determine the identity of mutations causing a phenotype. Here, we describe protocols for the construction of libraries for S. cerevisiae whole-genome sequencing. We also present a bioinformatic pipeline to determine the genetic variants in a yeast strain using whole-genome sequencing data. This pipeline can also be used for analyzing Schizosaccharomyces pombe mutants. © 2019 by John Wiley & Sons, Inc. Basic Protocol 1: Generation of haploid spores for bulk segregant analysis Basic Protocol 2: Extraction of genomic DNA from yeast cells Basic Protocol 3: Shearing of genomic DNA for library preparation Basic Protocol 4: Construction and amplification of DNA libraries Support Protocol 1: Annealing oligonucleotides for forming Y-adapters Support Protocol 2: Size selection and cleanup using SPRI beads Basic Protocol 5: Identification of genomic variants from sequencing data.