何首乌总RNA提取方法的比较及改进  

Comparison and Improvement of Methods for Total RNA Isolation from Polygonum multiflorum Thunb

作　　者：谭雪梅[1] 申彦晶[1] 严萍[1] 赵树进[2] 

机构地区：[1]华南理工大学生物科学与工程学院,广东广州510640 [2]广州军区广州总医院药学部,广东广州510010

出　　处：《生物技术通讯》2008年第3期414-416,共3页Letters in Biotechnology

基　　金：广东省科技厅项目(63108)

摘　　要：目的:探讨不同提取方法对提取何首乌总RNA的质量影响,寻找适于何首乌成熟叶组织RNA的提取方法。方法:以何首乌成熟叶组织为材料,采用SDS/酸酚法、常规CTAB法及改良的TRIzol试剂法分别进行实验,并对所提取RNA的质量进行验证。结果:采用3种方法都能提取出RNA,但质量差异较大。其中改良的TRIzol试剂法能有效抑制次生物质的影响,提取的RNA产量可达70～110μg/g,纯度高于其他2种方法,D260nm/D280nm值为1.85～1.97。结论:改良的TRIzol试剂法操作简便,提取的RNA完整性和纯度较高,可以满足下一步实验的要求。Objective： To approach an effective method for extracting total RNA from Polygonum multiflorum Thunb. Methods： By respectively employing SDS/acid phenol method, ordinary CTAB method or modified TRIzol method to extract total RNA from mature leaf tissues of P.multiflorum, moreover dectecting the quality of total RNA isolated by different methods. Results： The quality of total RNA isolated respectively by three methods were extremely different. Only the modified TRIzol method showed better integrity and purity of RNA, and efficiently inhibited the influence of carbohydrate, polyphenol and other secondary substances. The yield of total RNA isolated via this protocol could be obtained 70-110 μg per gram fresh tissues and the ratio of D260nm/D280nm were 1.85-1.97. Conclusion： RNA extracting with the modified TRIzol method can be absolutely used to RT-PCR or cDNA library construction.

关 键 词：何首乌 RNA提取 次生物质 RT-PCR 

分 类 号：Q522[生物学—生物化学]