| 野生型工业酿酒酵母Miseq测序方法的建立 |
| |
| 引用本文： | 曹德民,;张穗生,;罗贞贞,;黄日波.野生型工业酿酒酵母Miseq测序方法的建立[J].广西农业生物科学,2014(3):655-660. |
| |
| 作者姓名： | 曹德民 ;张穗生 ;罗贞贞 ;黄日波 |
| |
| 作者单位： | [1]广西大学生命科学与技术学院,南宁530003; [2]广西科学院广西生物炼制重点实验室,非粮生物质酶解国家重点实验室,国家非粮生物质能源工程技术研究中心,南宁530007 |
| |
| 基金项目： | 国家863课题（2012AA022106,2013AA050701）; 国家国际合作项目（2010DFB63590）; 广西科学研究和技术开发项目（桂科重12118004-2,桂科重1348004-1,桂科重1348004-3,桂科合1346011-4）; 广西自然科学基金（2011GNSFA-018113,2012AA022106）; 广西八桂学者建设工程专项经费; 广西科技创新能力与条件建设计划项目（桂科能12237022）; 广西科学基金项目13-051-08; 广西科学院基本科研业务费资助项目12YJ25SW03共同资助 |
| |
| 摘 要： | 新一代测序方法在基因组学研究应用日趋广泛,已经成为工业酿酒酵母菌株基因组学研究的重要技术平台,但对工业酿酒酵母新一代测序技术方法缺乏详细报道。Miseq是小型新一代基因组测序仪,本文报道我们自建的野生型工业酿酒酵母基因组Miseq测序方法。该方法包括：制备分离a型和α型交配型的单倍体菌株、采用电泳和分光光度法方法监控基因组文库建立、优化上机文库浓度后测序。其中电泳和分光光度法方法为首创的文库质量监控简易方法,质控检测得到酿酒酵母工业菌株测序条件为：菌株的基因组文库DNA片段大小为250-850bp且主要集中在350-550bp,文库浓度范围为7-13ng/μL;上机测序文库的优化浓度为20pM。
|
| 关 键 词： | 酿酒酵母 野生型工业菌株 Miseq 基因组测序方法 文库质量 |
Development of a Method to Sequence a Wild-type Industrial Strain of Saccharomyces cerevisea with Miseq |
| |
| Institution: | Cao Dernin , Zhang Suisheng , Luo Zhenzhen , Huang Ribo( 1 Life Science and Technology College, Guangxi University, Nanning, 530003; 2 Guangxi Key Laboratory of Biorefinery, State Key Laboratory of Non-food Biomass and Enzyme Technology, National Engineering Research Center for Non-food Biorefinery, Guangxi Academy of Science, Nanning, 530007) |
| |
| Abstract: | Next-generation DNA sequencing technology has become one of the most wilely used genomics tools and an important tools platform to investigate industrial yeast genome. However, there have not yet been any detail reports concerning how to sequence the genome of industrial S. cerevisea strain. Miseq is a bench-top next-generation sequencer. Here we reported an efficient self-design protocl of sequencing the genome of wild-type industrial S. cerevisea strain with Miseq. The key steps of the method include： Preparation of a and αtype haploid of the strain, confirmation of library quality for seqencing with electrophoresis and spectrometric analysis, sequcencing the genome with Miseq after the library concentration optimization. Our method first determines the genome library quality by electrophoresis and spectrometric analysis. We have found that DNA fragment of seqencing libary should be 250-850 bp, mainly in the 350-550 bp range and the concentration of the libray was 7-13 ng/μL, and the optimized concentration of library sequenced is 20 pM. |
| |
| Keywords: | Saccharomyces cerevisiae Wild-type industrial strain Miseq Genome sequencing method Library quality |
| 本文献已被 维普 等数据库收录！ |
|
