一种木本植物RNA和DNA的简捷提取方法

被引：5

作者：

黄真池

余秋梅

欧阳乐军

曾富华

机构：

[1] 湛江师范学院生命科学与技术学院

来源：

湖北农业科学 | 2010年 / 49卷 / 04期

基金：

广东省自然科学基金;

关键词：

木本植物; RNA; DNA; 非变性琼脂糖电泳;

D O I：

10.14088/j.cnki.issn0439-8114.2010.04.010

中图分类号：

Q943.2 [植物基因工程];

学科分类号：

摘要：

将尾叶桉(Eucalyptus urophylla)叶片在液氮中研磨成粉,先用含山梨醇、聚乙烯吡咯烷酮(PVP)、牛血清白蛋白、聚乙二醇和亚精胺的提取缓冲液使总核酸沉淀,同时去除了大部分次生代谢物质。然后用山梨醇、月桂酰肌氨酸钠进一步裂解细胞,释放核酸。再用CTAB与核酸结合成复合物溶于高盐溶液。用氯仿/异戊醇除蛋白质后,离心得含总核酸的上清液。运用钙盐分步沉淀法,先在上清液中加入1/10体积的10%氯化钙溶液,使DNA与Ca2+结合成DNA钙盐。向溶液中加入1/5体积的乙醇,使DNA钙盐形成沉淀析出,离心得DNA后,再增大乙醇浓度使RNA钙盐沉淀。得到的DNA和RNA用非变性琼脂糖凝胶进行质量检测,可得完整的RNA和DNA条带。此法经济、快捷,可同时得到DNA和RNA。

引用

页码：773 / 774+784 +784

页数：3

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