百合叶片总RNA提取方法比较及优化_尹慧.pdfVIP

中国农业大学学报 2008, 13( 4) :4 1245 Journal of China Agricultural University RNA 尹 慧 陈莉 李晓艳 陈秋明 义鸣放 ( 中国农业大学 农学与生物技术学院, 北京 1001 3 ) RNA , - . , CTAB、 SDS Trizol、CTAB、SDS RNA 。: SDS Trizol 、, CTAB RNA DNA 。 , CT AB , , / , DNase Ñ DNA, LiCl RNA 。RNA , D260/ D280 21 0, D260/ D230 21 1, 28S、18S , 11 5。RT2PCR , CTAB RNA 。 ; ; RNA ; CT AB ; ; RT2PCR S 6821 2 100724333(2008)042 004 12 05 A Analysis and improvement of high2quality RNA extraction in leaves of lily YIN Hui, CHEN Li, LI Xia o2yan, CHEN Qiu2ming , YI Ming2 fa ng ( College of Agriculture and Biotechnology, China Agricultural University Beijing 1001 3 China) Ab stra ct Five extraction methods: common Trizol, SDS/ phenol extraction, CTAB and the modified methods of SDS/ phenol extraction and CTAB were compared for extracting total RNA from lily leaves . The SDS and Trizolmethods were not effective in removing polysaccharides, while the RNA extracted by traditional CTAB method was ineffective in remo2 ving DNA. Because lily leaves are rich in polysaccharides and secondary metabolites, the modified CTAB method was improved by adding KAc and ethanol to remove polysaccharides, phenol/ chloroform to remove protein, LiCl to precipi2 tate RNA and DNase Ñto remove DNA. The modified CTAB can extract high2 quality and high2integrity RNA, as deter2 mined by UV2 spectroscopic analysis showing the D / D at 2. 0 and D / D at 2. 1. The electrophoresis profile 260 280 260 230 showed that the lightness at 28S was 11 5 times higher than that at 18S. The modified CTAB can efficiently remove poly2 saccharides, protein and DNA. The results