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相关概念视频

Genome Annotation and Assembly03:36

Genome Annotation and Assembly

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The genome refers to all of the genetic material in an organism. It can range from a few million base pairs in microbial cells to several billion base pairs in many eukaryotic organisms. Genome assembly refers to the process of taking the DNA sequencing data and putting it all back together in a correct order to create a close representation of the original genome. This is followed by the identification of functional elements on the newly assembled genome, a process called genome annotation.
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DNA isolation protocols can be fast and straightforward or complex and time-consuming depending on the type and quality of DNA required for further processing. For example, plasmid DNA extraction is a bit more complicated than genomic DNA extraction because of the need for an appropriate lysis method to separate plasmid DNA from gDNA during isolation. However, for specific applications, such as long-range DNA sequencing that require a good yield of high- quality DNA samples, we need to follow...
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Use of In Vivo Assembly for High-efficiency Plasmid Construction
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在活体DNA组装中使用PEDA方法.

Tianyuan Su1, Qingxiao Pang2, Qingsheng Qi3

  • 1State Key Laboratory of Microbial Technology, Shandong University, Qingdao, People's Republic of China.

Methods in molecular biology (Clifton, N.J.)
|March 12, 2024
PubMed
概括

菌体酶辅助的体内DNA组装 (PEDA) 能够在各种微生物中直接组装DNA部分. 这种高效的方法与体外技术相竞争,加速了合成生物学进步.

关键词:
组装DNA组件的DNA组件在活体中,在活体中.分子克隆是一种分子克隆.多种微生物多种微生物T4 DNA 连接酶的作用在T5DNA外核酶中.

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科学领域:

  • 合成生物学 合成生物学
  • 分子生物学分子生物学
  • 微生物学 微生物学

背景情况:

  • 对合成生物学来说,DNA组装至关重要.
  • 现有的方法可能很复杂或范围有限.
  • 需要高效,多功能的体内DNA组装技术.

研究的目的:

  • 引入和验证PEDA方法用于直接in vivoDNA组装.
  • 为了证明PEDA在各种微生物宿主中的适用性.
  • 为PEDA实施提供一个协议.

主要方法:

  • 菌体酶辅助的体内DNA组装 (PEDA) 协议.
  • 在大肠杆菌,Ralstonia eutropha,Pseudomonas putida,Lactobacillus plantarum和Yarrowia lipolytica中进行PEDA测试.
  • 使用短同源序列 (低至5bp) 组装DNA片段.

主要成果:

  • PEDA促进了DNA部分在体内直接组装.
  • 在多种细菌和酵母菌种中成功应用.
  • 实现了与体外方法相比较的高组装效率.
  • 经过证明的组装,具有最小的同质手臂 (5 bp).

结论:

  • PEDA是一种简单,高效和多功能工具,用于体内DNA组装.
  • 该方法扩大了合成生物学研究的工具包.
  • 在各种微生物中,PEDA支持DNA分子的快速构建.