Richards E, Reichardt M, Rogers S
Washington University, St. Louis, Missouri, USA.
Curr Protoc Mol Biol. 2001 May;Chapter 2:Unit2.3. doi: 10.1002/0471142727.mb0203s27.
This unit describes two methods for preparing genomic DNA from plant tissue. In the first method, plant cells are lysed with ionic detergent, treated with protease, and subsequently purified by cesium chloride (CsCl) density gradient centrifugation. The second method is based upon a series of treatments with the nonionic detergent cetyltrimethylammonium bromide (CTAB) to lyse cells and purify nucleic acid. Nucleic acid is recovered from the final CTAB solution by isopropanol or ethanol precipitation. The first method, although somewhat more lengthy, results in highly purified nucleic acid. The second method requires fewer manipulations, results in very high yields (approximately 10-fold higher per gram fresh tissue depending on species and condition of starting material), and produces DNA that is less pure but nonetheless suitable in quality for use in many molecular biology manipulations.
本单元介绍了两种从植物组织中制备基因组DNA的方法。第一种方法是用离子去污剂裂解植物细胞，用蛋白酶处理，随后通过氯化铯（CsCl）密度梯度离心进行纯化。第二种方法基于用非离子去污剂十六烷基三甲基溴化铵（CTAB）进行一系列处理来裂解细胞并纯化核酸。通过异丙醇或乙醇沉淀从最终的CTAB溶液中回收核酸。第一种方法虽然稍微冗长一些，但能得到高度纯化的核酸。第二种方法所需的操作较少，产量非常高（根据起始材料的物种和状况，每克新鲜组织的产量大约高10倍），并且产生的DNA纯度较低，但质量仍适用于许多分子生物学操作。
