木薯块根总RNA提取方法的比较和改进

[目的]为更好地利用基因操作手段改良木薯品质和提取高质量的木薯总RNA提供科学依据。[方法]采用4种方法提取木薯块根RNA,筛选一种最简单、有效的方法,并利用RT-PCR技术克隆木薯淀粉分支酶,分析所提取RNA的质量。[结果]结果表明,用改进的CTAB法和plant RNAReagent kit均能提取高质量的RNA,用plant RNAReagent更能节省时间,而且纯度比较高。对plant RNAReagent提取的RNA进行RT-PCR克隆,克隆出了与木薯淀粉合成有关的SBEII基因。[结论]plant RNAReagent kit提取的RNA,其质量可以满足基本的试验要求。

Comparison and Improvement of the Method of Extraction Total RNA form Tubers of Manihot esculenta Crantz

[Objective] The purpose was to provide the scientific basis for improving the quality of Manihot esculenta Crantz and extracting the high-quality total RNA form tubers of Manihot esculenta Crantz by genic method.[Method]The best simple and effective method from four measures which can extract RNA form tubers of Manihot esculenta Crantz was applied.Branching enzyme of cassava was cloned with RT-PCR technology and the quality of RNA extracted was analyzed.[Result] The results showed that the developed CTAB and plant RNA Reagent kit could get the high-quality RNA,and the latter was with the less time and higher purity.By RT-PCR cloning RNA extracted with plant RNA Reagent kit,SBELL gene which was related to synthesis of the cassava was got.[Conclusion]The quality of RNA extracted with plant RNA Reagent kit can meet the demands for experiment.