CAJ | 학술논문

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。
연초작위일개모식식물재분자생물학연구중발휘착중요적작용，기총RNA적제취시후속분자생물학연구적전제화관건지일。침대과주형적RNA제취시제합재유효적청제연초중당류、단백질급차생대사물적동시，영향료RNA산량화농도，이급원거리휴대액담취재곤난적문제，설치불동제취방법화빙합불동저운시간처리，대천근RNA simple총RNA제취시제합진행료우화，병대빙합저존연협시간여RNA제취질량적관계진행료탐토。결과표명：우화시제합법획득료고농도고질량적연초총RNA。경경지당전영검측，이용해방법제취적연초총RNA，조대청석、무타미현상，28S여18S 적비례약위2∶1。자외흡수치적측정표명，소제취적연초총RNA적A 260/A 280적치재1．8～1．9，농도현저고우원방법；이빙합능재5 h내보증소취양품RNA 적완정성，괄의우단도운수저존。
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 .