低温切片破碎紫花地丁根组织提取RNA的研究  

Research the effective and simple Methods for Breaking the root of the Herb Violae to Extract RNA

作　　者：毛樱逾[1] 罗波[1] 罗茂[1] 段素群[1] 张春[1] 

机构地区：[1]泸州医学院,四川泸州646000

出　　处：《时珍国医国药》2013年第11期2816-2818,共3页Lishizhen Medicine and Materia Medica Research

基　　金：国家自然科学基金(No.81001700);四川省教育厅青年基金项目(No.11ZB227);四川省科技厅应用基础计划(No.2012JY0081)

摘　　要：目的通过比较液氮研磨法和低温切片法破碎紫花地丁根组织来提取RNA的效果,以获得提取药用植物紫花地丁根RNA的最佳方法。方法用液氮研磨法和低温切片法破碎紫花地丁根组织,然后提取RNA。通过RNA浓度、纯度及完整性的检测,以及扩增GAPDH基因来比较两种方法的效果。结果液氮研磨法和低温切片法提取的RNA浓度分别为1.21,3.57μg/μl。琼脂糖凝胶电泳检测可见两条明显的条带。28S rRNA与18S rRNA条带亮度之比大于1。GAPDH基因的PCR扩增在约230 bp处有与预期长度一致的明亮条带。结论低温切片法破碎植物根组织能达到液氮研磨法的效果,是提取RNA时,破碎植物根组织的一种成本低,效果好,快速简便的方法。Objective Comparing liquid nitrogen grinding method with frozen section method for breaking the root of the herb Violae to extract RNA, to demonstrate that frozen section is a simple and effective mathod. Methods Broke the root of the Herb Violae by liquid nitrogen grinding method and frozen section method, and extracted RNA. Concentration, purity and integrity of RNA were detected. The GAPDH gene was amplified by RT - PC R. Results The RNA concentration was 1.21 μg/μl and 3.57 μg/μl, extracted by liquid nitrogen grinding method and frozen section method respectively. Two distinct bands were showed by agarose gel electro- phoresis. The ratio of brightness of the 28S rRNA and the 18S rRNA bands was greater than 1. The length of GAPDH gene was 230 bp. Conclusion The experimental results indicated that frozen section method to break the root of the Herb Violae can meet the needs of most molecular biological experiments including gene cloning and expression analysis. It is an effective and simple method for extract RNA of from the plant root.

关 键 词：低温切片 RNA提取 紫花地丁根 

分 类 号：R2-03[医药卫生—中医学]