生物技术通报 ›› 2016, Vol. 32 ›› Issue (7): 34-39.doi: 10.13560/j.cnki.biotech.bull.1985.2016.07.005

• 技术与方法 • 上一篇    下一篇

高山被孢霉的基因组及转录组的提取技术研究

凌瑞1, 孙立洁1, 2, 陈祥松1, 袁丽霞1, 吴金勇1, 姚建铭1   

  1. 1. 中国科学院合肥物质科学研究院等离子体物理研究所,合肥 230031;
    2. 中国科学院湖北产业技术创新与育成中心,武汉 430072
  • 收稿日期:2015-09-06 出版日期:2016-07-25 发布日期:2016-07-25
  • 作者简介:凌瑞,女,硕士研究生,研究方向:微生物发酵工程与代谢调控;E-mail:lrr@mail.ustc.edu.cn
  • 基金资助:
    国家高技术研究发展计划(“863”计划)(2014AA021703),教育部留学回国人员科研启动基金资助项目(教外司留[2013]693号)

Technologies of Extracting Genome and Transcriptome from Mortierella alpina

LING Rui1, SUN Li-jie1, 2, CHEN Xiang-song1, YUAN Li-xia1, WU Jin-yong1, YAO Jian-ming1   

  1. 1. Institute of Plasma Physics Chinese Academy of Sciences,Hefei Institute of Physical Science,Chinese Academy of Sciences,Hefei 230031;
    2. Hubei High-tech Innovation and Business Incubation Center,Chinese Academy of Sciences,Wuhan 430072
  • Received:2015-09-06 Published:2016-07-25 Online:2016-07-25

摘要: 基因组和转录组的高通量测序对样品的纯度和含量要求较高,旨在通过研究高山被孢霉在不同培养条件下和采用不同菌丝处理方法对其基因组和转录组质量的影响来确定最佳提取方式。结果表明,PDA培养基培养的菌丝与发酵培养基培养的菌丝经基因组提取后在琼脂糖凝胶电泳中有明显不同的表现,使用发酵培养基培养的菌丝提取的基因组DNA无降解,而以PDA培养基培养的菌丝提取的基因组DNA出现部分降解。在使用国产试剂盒做高山被孢霉转录组RNA提取实验中发现,使用液氮速冻后破碎菌丝并结合使用β-疏基乙醇更易提取到高浓度且无降解的转录组样品。

关键词: 高山被孢霉, 基因组, 转录组, 提取技术

Abstract: High-throughput sequencing of transcriptome and genome demands the high purity and content of sample,the purpose of this work is to determine the optimal extraction technology by studying the effects of Mortierella alpina in different culture conditions and using different hypha processing methods on the genome and transcriptome quality. The results showed that the mycelium from liquid fermentation medium after genome extraction presented a significant difference in agarose gel electrophoresis compared to the mycelium from PDA culture medium,the former gave degradation-free genome DNA but the latter was extracted genomic DNA of some degradation. During using domestic brand kit to study M. alpina tanscriptome RNA extraction,it was found that using liquid nitrogen fast-frozen method to break the hypha cell wall combined with applying β-Mercaptoethanol resulted in more likely to extract the high concentration and no-degradation transcription sample.

Key words: Mortierella alpina, genome, transcriptome, extraction technology