激光生物学报2011，Vol.20Issue(1)：108-115,8.DOI:10.3969/j.issn.1007-7146.2011.01.021
茶树不同器官组织总RNA提取方法的研究
A Simple1 and Effective Method for Extracting Total RNA from Different Organs of Tea Plant [Camellia sinensis (L.) O. Kuntze]
摘要
Abstract
Obtaining high quality total RNA is an important prerequisite for studying genomics and functional genomics of tea plant. It is hard to extract high quality total RNA from tea due to the interference of RNase, and polyphenols. Aiming at the drawbacks in extracting method for total RNA of tea plant,a comparative study was carried out to determine material dosage, extracting buffer, protein extracting solution,precipitating reagent for extracting total RNA, in this paper. The simple CTAB-LiCl method was established, which was a simple and effective method for extracting total RNA from different organs of tea plant. The differences among the improved Tri-Reagent method, the improved CTAB method and the simple CTAB-LiC1 method were comparably investigated. The results of spectrophotometry testing and agarose gel electrophoresis showed that the simple CTAB-LiC1 method was suitable for extracting high yield and high quality total RNA from tea organs. The intensity ratio of 28 S and 18 S rRNA bands was 2:1, the absorbance ratios of A260 to A280 were about 2.0, the absorbance ratios of A260 to A230 were up to 2.0, and the yield of total RNA ranged from 290.61 μg/g to 532.44 μg/g fresh weight. This method is good for extracting total RNA from different tea organs with its simple, efficient, high extracting rate and low cost. The results of RT-PCR and cDNA-AFLP analysis also revealed that the simple CTAB-LiC1 method could obtain high quality total RNA from different tea organs,关键词
茶树/总RNA/器官/提取方法/简易CTAB-LiC1法Key words
tea plant (Camellia sinensis (L.) O. Kuntze)/ total RNA/ organs/extracting method/ CTAB-LiCl method分类
生物科学引用本文复制引用
杨冬青,王云生,孙美莲,单育,骆洋,韦朝领,高丽萍,夏涛..茶树不同器官组织总RNA提取方法的研究[J].激光生物学报,2011,20(1):108-115,8.基金项目
国家"973"计划前期项目(2007CB116211) （2007CB116211）
国家自然科学基金项目(30771755,30972401) （30771755,30972401）
安徽省自然科学基金项目(090411006) （090411006）
