Schenk John J, Becklund L Ellie, Carey S James, Fabre Paige P
Department of Environmental and Plant Biology Ohio University Athens Ohio 45701-2979 USA.
Appl Plant Sci. 2023 Jun 2;11(3):e11517. doi: 10.1002/aps3.11517. eCollection 2023 May-Jun.
Cetyltrimethylammonium bromide (CTAB)-based methods are widely used to isolate DNA from plant tissues, but the unique chemical composition of secondary metabolites among plant species has necessitated optimization. Research articles often cite a "modified" CTAB protocol without explicitly stating how the protocol had been altered, creating non-reproducible studies. Furthermore, the various modifications that have been applied to the CTAB protocol have not been rigorously reviewed and doing so could reveal optimization strategies across study systems. We surveyed the literature for modified CTAB protocols used for the isolation of plant DNA. We found that every stage of the CTAB protocol has been modified, and we summarized those modifications to provide recommendations for extraction optimization. Future genomic studies will rely on optimized CTAB protocols. Our review of the modifications that have been used, as well as the protocols we provide here, could better standardize DNA extractions, allowing for repeatable and transparent studies.
基于十六烷基三甲基溴化铵（CTAB）的方法被广泛用于从植物组织中分离DNA，但植物物种间次生代谢产物独特的化学组成使得有必要对该方法进行优化。研究文章常常引用“改良的”CTAB方案，却未明确说明该方案是如何改变的，从而导致研究无法重复。此外，应用于CTAB方案的各种改良方法尚未经过严格审查，而这样做可能会揭示跨研究系统的优化策略。我们查阅了用于分离植物DNA的改良CTAB方案的文献。我们发现CTAB方案的每个阶段都已被修改，并且我们总结了这些修改内容以提供提取优化的建议。未来的基因组研究将依赖于优化的CTAB方案。我们对已使用的改良方法以及此处提供的方案进行的综述，能够更好地规范DNA提取，从而实现可重复且透明的研究。
