烟草RNA提取方法的优化及冰盒储存时间的探讨

The Optimization of RNA Isolation and the Exploration of Ice Box Storage Time in Tobacco

烟草作为一个模式植物在分子生物学研究中发挥着重要的作用，其总RNA的提取是后续分子生物学研究的前提和关键之一。针对过柱型的RNA提取试剂盒在有效的清除烟草中糖类、蛋白质及次生代谢物的同时，影响了RNA产量和浓度，以及远距离携带液氮取材困难的问题，设置不同提取方法和冰盒不同储运时间处理，对天根RNA simple总RNA提取试剂盒进行了优化，并对冰盒贮存烟叶时间与RNA提取质量的关系进行了探讨。结果表明：优化试剂盒法获得了高浓度高质量的烟草总RNA。经琼脂糖电泳检测，利用该方法提取的烟草总RNA，条带清晰、无拖尾现象，28S与18S 的比例约为2∶1。紫外吸收值的测定表明，所提取的烟草总RNA的A 260/A 280的值在1．8～1．9，浓度显著高于原方法；而冰盒能在5 h内保证所取样品RNA 的完整性，适宜于短途运输储存。

As a model plant ,tobacco plays an important role in molecular biology research ,the total RNA extraction is one of the key prerequisites in a subsequent molecular biology .In view of the problem that colum-nar RNA isolation kits effectively remove tobacco carbohydrates ,proteins and secondary metabolites but at the same time impact of RNA yield and concentration ,and that long-distance carriage of liquid nitrogen is diffi-cult,different extraction methods and different storage time to deal with ice box were set ,the TIANGEN RNA simple total RNA extraction kit was optimized, and the relationship between ice box storage time and RNA ex-traction quality of tobacco was discussed .The results showed that：the improved method obtained a high concen-tration and high-quality tobacco total RNA .By agarose gel electrophoresis ,the tobacco RNA extracted with this method had clear bands ,no smearing ,the ratio of 28S to 18S was approximately 2∶1.UV absorption value de-termination showed that the ratio ofA 260/A 280 was in the range of 1.8-1.9,and the concentration was signifi-cantly higher than that with the original method;while the ice box could guarantee the integrity of the RNA samples in5 hours ,suitable for short-distance transport storage .