High-Quality Genomic DNA Extraction Protocol for Bacillus and Clostridium Species

Current protocols Pub Date : 2024-10-07 DOI:10.1002/cpz1.70027

Yohannes Beyene Mekonnen, Marina Elisabeth Aspholm, Ephrem Debebe Zegeye

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Abstract

High-quality DNA with sufficient yield is the goal of DNA extraction protocols. We present an optimized, cost-effective method for extracting next-generation sequencing (NGS)-quality genomic DNA from Bacillus and Clostridium species using the chloroform-isoamyl approach. The protocol involves two main procedures: cultivation of the bacteria under appropriate conditions, followed by DNA extraction through cell lysis, phase separation, DNA precipitation, and cleanup. This method has been successfully applied to several hundred strains of Bacillus and Clostridium species in our laboratory, including B. cereus, B. licheniformis, C. sporogenes, and C. tyrobutyricum, demonstrating its efficacy and reliability for producing high-quality DNA that meets NGS quality control standards. © 2024 The Author(s). Current Protocols published by Wiley Periodicals LLC.

Basic Protocol 1: Culturing Bacillus and Clostridium species

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芽孢杆菌和梭状芽孢杆菌的高质量基因组 DNA 提取规程。

具有足够产量的高质量 DNA 是 DNA 提取方案的目标。我们提出了一种优化的、经济有效的方法，利用氯仿-异戊基法从芽孢杆菌和梭状芽孢杆菌中提取下一代测序（NGS）质量的基因组 DNA。该方法包括两个主要步骤：在适当条件下培养细菌，然后通过细胞裂解、相分离、DNA 沉淀和净化提取 DNA。在我们的实验室中，这种方法已成功应用于数百株芽孢杆菌和梭状芽孢杆菌，包括蜡样芽孢杆菌、地衣芽孢杆菌、孢子芽孢杆菌和酪酸梭状芽孢杆菌，证明了它在产生符合 NGS 质量控制标准的高质量 DNA 方面的有效性和可靠性。© 2024 作者。当前协议》由 Wiley Periodicals LLC 出版。基本规程 1：培养芽孢杆菌和梭状芽孢杆菌基本规程 2：DNA 提取 © 2024 by John Wiley & Sons, Inc.

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Current protocols

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