使用重复的溶解步骤,从异性蓝菌培养中提取的异性蓝菌和蓝菌DNA之间进行分离
Alexis D Wagner1, Mohammed M A Ahmed2, Victoria Starks1
1Department of BioMolecular Science, School of Pharmacy, University of Mississippi, University, Mississippi, 38677, United States of America.
bioRxiv : the preprint server for biology
|September 4, 2024
概括
这项研究通过使用代溶解和尺寸选择步骤优化了从蓝藻细菌中提取DNA的过程. 这种方法成功丰富了宿主DNA,使两种蓝藻细菌物种的完整基因组测序成为可能.
科学领域:
- 微生物学 微生物学
- 基因组学就是基因组学.
- 分子生物学分子生物学
背景情况:
- 菌DNA提取具有挑战性,因为细胞结构多样化和常见的异质型微生物群.
- 低的蓝藻细菌DNA产量和来自其他物种的高污染阻碍了完整基因组测序.
研究的目的:
- 优化DNA分离协议,以从混合培养中丰富蓝色细菌宿主DNA.
- 通过纳米孔技术实现蓝藻细菌的成功基因组测序.
主要方法:
- 开发了一种采用三个代细胞溶解步骤的协议,以减少微生物组DNA.
- 整合了两个基于珠子的大小选择步骤,以删除低分子量DNA片段.
- 利用EPI2ME分析来评估异质型微生物组的减少.
主要成果:
- 代溶解显著降低了异质微生物组DNA的比例,每一步.
- 优化的协议产生了适合纳米孔测序的DNA,克服了剪切挑战.
- 成功测序并组装了 *Leptolyngbya* sp. 的封闭染色体. BL-A-14 (7.2 Mb) 和 *Limnothrix* sp. 的使用情况. 在BL-A-16 (4.5 Mb).
结论:
- 代溶解和尺寸选择协议有效地丰富了蓝菌DNA进行测序.
- 这种方法对于测序异性菌培养有价值,在异性菌培养是不可行的情况下.
- 该方法还为研究相关的异质型微生物组组成提供了潜力.
相关概念视频
DNA Isolation
38.5K
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...
38.5K
DNA Agarose Gel Electrophoresis
96.3K
Agarose gel electrophoresis is a laboratory technique commonly used to separate DNA fragments by size. However, it can also be used to isolate and purify DNA fragments using a gel extraction protocol.
Gel extraction follows five major steps: running gel electrophoresis to separate fragments, isolating the individual bands, extracting DNA from those bands, and removing the dye and salts from the extracted mixture to obtain pure DNA.
In cloning experiments, both the insert and vector DNA...
Gel extraction follows five major steps: running gel electrophoresis to separate fragments, isolating the individual bands, extracting DNA from those bands, and removing the dye and salts from the extracted mixture to obtain pure DNA.
In cloning experiments, both the insert and vector DNA...
96.3K


