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在CTAB中优化溶解步骤,提取二氧化干燥和草本树叶组织的DNA

S James Carey¹, L Ellie Becklund¹, Paige P Fabre¹

¹Department of Environmental and Plant Biology Ohio University Athens Ohio 45701-2979 USA.

Applications in plant sciences

|June 21, 2023

概括

使用 cetyltrimethylammonium bromide (CTAB) 优化植物DNA提取需要调整溶解条件. 凝保存和特异性缓冲产生更高的DNA质量和数量,改善研究结果.

科学领域:

分子生物学分子生物学
植物科学植物科学
生物化学生物化学

背景情况:

乙三甲基化物 (CTAB) 是一种具有成本效益的植物DNA提取方法.
现有的CTAB协议往往是经验性的修改,缺乏对DNA产量和完整性单个参数效应的系统评估.

研究的目的:

系统地研究化学添加剂,化温度和溶解时间对植物DNA数量和质量的影响.
确定基于CTAB的DNA提取的最佳条件,以提高产量和纯度.

主要方法:

评估了各种化学添加剂 (例如,聚烯利),化温度和溶解时间对DNA提取的影响.
比较DNA产量,碎片长度和纯度从凝保存与草药馆保存的植物组织.
利用CTAB和修改的CTAB缓冲器进行DNA隔离.

主要成果:

化温度和溶解持续时间影响了DNA度和碎片长度,但提取剂的纯度受到最显著的影响.
带有聚烯罗立缓冲体的CTAB和CTAB产生了最高的DNA质量和数量.
与草药组织相比,凝保存的组织产生了显著更高的DNA产量,更长的DNA片段和更纯的提取剂.

结论:

建议使用凝保存的组织进行DNA提取.
建议采用较短,较冷的溶解步骤用于凝保存的组织,以提高纯度,防止DNA碎片化,减少处理时间.

关键词:

在CTABB中,它是最重要的.提取DNA 提取DNA的方法细胞溶解细胞的溶解.乙烯基三甲基氨酸化物分子生物学分子生物学植物遗传学植物遗传学

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