摘要
本实验利用TRIZOL试剂提取水稻苗期叶片总RNA,对三种常用的RNA纯化方法和改进的纯化方法进行了比较。通过凝胶电泳、紫外分光光度法和RT-PCR法检测提取的RNA样品的品质,结果表明,应用改进的RNA纯化方法可有效去除水稻RNA中的DNA污染,电泳条带清晰无降解;OD260/OD280为1.91~1.98,具有较高的纯度;改良方法纯化的RNA逆转录为cDNA,作为PCR扩增的模板,以不同的循环数进行扩增,在反应适宜循环数内检测不到DNA片段污染,说明用改进方法提取的RNA,其质量和纯度可以满足下一步分子生物学研究的需要。
Total RNA was extracted from rice leaves using TRIZOL regent.Three commonly used RNA purification methods and an improved method were compared in this experiment.The quality of total RNA was analyzed through gel electrophoresis,UV spectrophotometry and RT-PCR.The RNA purified by the improved method gave clear bands of rRNA in the electrophoresis without DNA contamination and degradation.OD260/OD280 value was from 1.91 to 1.98.RNA purified by the improved method was used for RT-PCR and none band of DNA conta...
出处
《中国农业科技导报》
CAS
CSCD
2008年第S1期111-115,共5页
Journal of Agricultural Science and Technology
基金
国家973计划项目"农业转基因生物安全风险评价与控制基础研究"(2007CB109201)资助