预隔离分子选用于从环境中取出细菌微生物的高通量采样和测序
Soon Keong Wee1, Mark Boon Pho Chan1, Suppiah Paramalingam Sivalingam1
1Lee Kong Chian School of Medicine, Nanyang Technological University, Singapore.
Current protocols
|May 29, 2023
概括
这项研究为隔离和表征环境微生物提供了全面的协议. 它详细介绍了采样,丰富,查和全基因组测序的方法,以准确识别微生物和进行功能分析.
科学领域:
- 微生物学 微生物学
- 环境科学 环境科学
- 基因组学就是基因组学.
背景情况:
- 环境研究需要微生物培养和表征来理解生态作用.
- 纯培养能够对微生物进行详细的表型和功能分析.
- 为了有效地隔离罕见的微生物,通常需要进行丰富和PCR查.
研究的目的:
- 概述从环境样本中选,隔离和测序微生物的端到端协议.
- 为环境研究设计,丰富,选和隔离提供系统方法.
- 详细介绍物种识别和全基因组测序的技术.
主要方法:
- 环境样本采集 (土壤和水) 和微生物丰富.
- 使用定量PCR (qPCR) 进行查,并通过MALDI-TOF MS或桑格测序进行物种识别.
- 用牛津纳米孔平台进行基因组DNA提取和全基因组测序.
主要成果:
- 建立了从各种环境矩阵中对微生物隔离和表征的系统方法.
- 证明了 qPCR 和 MALDI-TOF MS 在物种鉴定中的有用性.
- 成功地应用了牛津纳米孔测序来对分离物进行全面的基因组分析.
结论:
- 提出的协议有助于有效和准确的环境微生物的选,隔离和基因组特征.
- 这些方法对于了解微生物的流行,分布,持久性和生态系统中的功能至关重要.
- 获得的综合基因组数据有助于为生态和功能研究进行菌株类型和基因型定型.
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