| 一种从富含多酚多糖的西葫芦组织中快速提取RNA的方法 |
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| 引用本文： | 周腾飞,伍林,秦晓蓉,童琨,童玲,胡适,秦悦,易德莲,刘郭飞,李丹.一种从富含多酚多糖的西葫芦组织中快速提取RNA的方法[J].湖北化工,2014(1):40-42. |
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| 作者姓名： | 周腾飞 伍林 秦晓蓉 童琨 童玲 胡适 秦悦 易德莲 刘郭飞 李丹 |
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| 作者单位： | [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 |
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| 基金项目： | 武汉科技大学绿色刺造和节能减排科研中心开放基金资助项目（B1207） |
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| 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．A250／280值为2.08、A250/280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。
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| 关 键 词： | CTAB 西葫芦 RNA提取 多糖 多酚 |
An Efficient Method for Extracting RNA from Cucurbita Pepo Tissues Rich in Polysaccharides and Polyphenols |
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| ZHOU Teng-fei,WU Lin,QIN Xiao-rong,TONG Kun,TONG Ling,HU Shi,QIN Yue,YI De-lian,LIU Guo-fei,LI Dan.An Efficient Method for Extracting RNA from Cucurbita Pepo Tissues Rich in Polysaccharides and Polyphenols[J].Hubei Chemical Industry,2014(1):40-42. |
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| Authors: | ZHOU Teng-fei WU Lin QIN Xiao-rong TONG Kun TONG Ling HU Shi QIN Yue YI De-lian LIU Guo-fei LI Dan |
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| Affiliation: | 1. Institute of Applied Chemistry ,Wuhan University of Science and Technology,Wuhan 430081 ,China ; 2. School of Life Sciences ,Wuchang University of Technology ,Wuhan 430223,China) |
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| Abstract: | 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. |
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| Keywords: | CTAB Cucurbita pepo RNA extraction polysaccharide polyphenol |
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