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A fully automatable enzymatic method for DNA extraction from plant tissues

机译:从植物组织中提取DNA的全自动酶法

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Background DNA extraction from plant tissues, unlike DNA isolation from mammalian tissues, remains difficult due to the presence of a rigid cell wall around the plant cells. Currently used methods inevitably require a laborious mechanical grinding step, necessary to disrupt the cell wall for the release of DNA. Results Using a cocktail of different carbohydrases, a method was developed that enables a complete digestion of the plant cell walls and subsequent DNA release. Optimized conditions for the digestion reaction minimize DNA shearing and digestion, and maximize DNA release from the plant cell. The method gave good results in 125 of the 156 tested species. Conclusion In combination with conventional DNA isolation techniques, the new enzymatic method allows to obtain high-yield, high-molecular weight DNA, which can be used for many applications, including genome characterization by AFLP, RAPD and SSR. Automation of the protocol (from leaf disks to DNA) is possible with existing workstations.
机译:与从哺乳动物组织中分离DNA不同,从植物组织中提取背景DNA仍然很困难,因为植物细胞周围存在刚性细胞壁。当前使用的方法不可避免地需要费力的机械研磨步骤,这对于破坏细胞壁释放DNA是必需的。结果使用不同碳水化合物酶的混合物,开发了一种方法,该方法能够完全消化植物细胞壁并随后释放DNA。消化反应的最佳条件使DNA剪切和消化减至最小,并使DNA从植物细胞中释放最大化。该方法在156个测试物种中的125个中获得了良好的结果。结论与传统的DNA分离技术相结合,新的酶促方法能够获得高产量,高分子量的DNA,可用于许多应用,包括通过AFLP,RAPD和SSR进行基因组鉴定。现有工作站可以实现协议(从叶盘到DNA)的自动化。

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