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 Ahmed1,2, Victoria A Starks1
1Department of BioMolecular Science, School of Pharmacy, University of Mississippi: Faser Hall, University, Mississippi, 38677, United States of America.
G3 (Bethesda, Md.)
|June 11, 2025
概括
使用代溶解和大小选择优化了对蓝藻细菌的DNA提取,显著增加宿主DNA比例,以改善基因组组装. 这种方法有助于测序异性蓝菌培养物.
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提取是具有挑战性的,因为不同的细胞结构和共发生的微生物.
- 在分离物中低的蓝藻细菌DNA比例阻碍了完整的基因组组装.
研究的目的:
- 优化DNA分离协议,用于从异种培养中丰富蓝菌DNA.
- 使用纳米孔技术实现蓝藻细菌的高质量基因组测序.
主要方法:
- 代细胞溶解 (三个步骤) 以提高蓝色细菌DNA产量.
- 基于珠子的尺寸选择,以减轻纳米孔测序的溶解中的DNA剪切.
- 分离DNA的纳米孔测序和生物信息分析.
主要成果:
- 代溶解使菌DNA的比例从35%增加到75%以上.
- 成功测序并组装了两个淡水蓝藻细菌 (Leptolyngbya sp. 在BL-A-14和Limnothrix sp.中使用. BL-A-16) 的一个部分.
- 在各自的分离物中,获得了7.2 Mb和4.5 Mb的基因组大小.
结论:
- 优化的协议有效地丰富了混合培养中的蓝藻细菌DNA.
- 这种方法促进了蓝藻细菌的基因组测序,即使没有菌培养.
- 这种方法对于在环境样本中研究多样化的蓝菌种群是有价值的.
更多相关视频
08:14Extraction of Non-Protein Amino Acids from Cyanobacteria for Liquid Chromatography-Tandem Mass Spectrometry Analysis
Published on: December 9, 2022
3.7K
10:03Isolation, Propagation, and Identification of Bacterial Species with Hydrocarbon Metabolizing Properties from Aquatic Habitats
Published on: December 7, 2021
4.3K
相关概念视频
DNA Isolation
38.4K
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...
38.4K
Tissue Homogenization and Cell Lysis
7.4K
Tissue homogenization involves disintegrating tissue architecture and lysing cells, and is an early step in isolating and analyzing cellular components. The method used for homogenization depends on the sample type, the amount of sample available, the analyte to be obtained, and the sensitivity of the method. These methods are broadly classified as mechanical and non-mechanical methods.
Mechanical methods of tissue homogenization
These methods rely on applying external physical force to disrupt...
Mechanical methods of tissue homogenization
These methods rely on applying external physical force to disrupt...
7.4K
