一种从富含多酚多糖的西葫芦组织中快速提取RNA的方法  

An Efficient Method for Extracting RNA from Cucurbita Pepo Tissues Rich in Polysaccharides and Polyphenols

作　　者：周腾飞[1] 伍林[1] 秦晓蓉[1] 童琨[1] 童玲[2] 胡适[1] 秦悦[1] 易德莲[1] 刘郭飞[1] 李丹[1] 

机构地区：[1]武汉科技大学应用化学研究所,湖北武汉430081 [2]武昌理工学院生命科学学院,湖北武汉430223

出　　处：《化学与生物工程》2014年第1期40-42,共3页Chemistry & Bioengineering

基　　金：武汉科技大学绿色制造和节能减排科研中心开放基金资助项目(B1207)

摘　　要：对比研究了一步法、尿素/氯化锂法、热酚法、LUG法等4种方法,建立了一种适合西葫芦组织RNA提取的改进的CTAB方法。用该方法从西葫芦叶中提取的RNA在琼脂糖凝胶电泳上清楚显示出28S和18S两条链,A260/280值为2.08、A260/230值为2.0,表明提取的RNA没有多酚、多糖、蛋白质污染;RNA产量为210μg·g-1鲜质量;对RT-PCR的结果进行测序,通过NCBI-BLAST比对可知,与印度南瓜的同源性达89%。用该方法从西葫芦果肉中也获取了高质量的RNA。改进的CTAB法全程只需2h,不用液氮、亚精胺、二硫苏糖醇、蛋白酶K等试剂。A comparative study on four kinds of methods for RNA extraction from Cucurbita pepo ,one-step method, urea/LiC1 method, hot phene method and LUG method was carried out, then an improved cetyltrimeth- ylammonium bromide （CTAB）-based RNA extraction method was developed. This method clearly showed 28S and 18S ribosomal RNA bands and produced RNA with high yield （210 μg·g^-1fresh mass） and high quality （A260/280 value 9.08;A260/280 value 2.0）,which indicated that the RNA was of high purity without polyphenol, polysaccharide and protein contamination. The result of RT-PCR was sequenced. Sequence alignment using the GenBank database revealed a high level of homology with Cucurbita maxima. Time for isolation of RNA using this method was 2 h. This method avoided the use of liquid nitrogen,spermidine,dithiothreitot （DTT）,protein- ase K which were unusual reagents in the laboratory.

关 键 词：CTAB 西葫芦 RNA提取 多糖 多酚 

分 类 号：O522[理学—高压高温物理]