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新型试剂-D.S.G在烟草RNA提取与纯化中的应用
引用本文:李正风 李文正 夏玉珍. 新型试剂-D.S.G在烟草RNA提取与纯化中的应用[J]. 勤云标准版测试, 2007, 0(3)
作者姓名:李正风 李文正 夏玉珍
作者单位:云南省烟草科学研究所,云南农业大学 云南昆明650201,云南省烟草科学研究所,红塔集团,云南玉溪653100,云南玉溪653100,云南玉溪653100
基金项目:国家自然科学基金项目“烟草富含甘氨酸RNA结合蛋白分子生物学研究”(30560062);中国烟草云南省公司科技开发计划项目“云南烟草基因组BAC文库的构建”(03A01):中国烟草云南省公司科技开发计划项目“烟草品质相关基因表达谱分析及在育种实践中的应用”(05-15).
摘    要:核酸提取是分子生物学实验的基础,提取高质量的核糖核酸(RNA)是构建cDNA文库和进行基因表达研究工作的前提。笔者比较了TRIREAGEN法、D.S.G法、异硫氰酸胍氯化铯密度梯度分离法、D.S.G和TRIREAGENT结合法、TRIREAGENT和D.S.G结合法等5种烟草RNA提取方法,结果表明D.S.G和TRIREAGENT两种试剂的配合使用均可有效地去除烟草植物细胞壁中的多聚糖及其它成份,获取较高质量的RNA。这说明新型试剂-D.S.G与TRIREAGENT的配合使用有利于获得高质量的RNA,但D.S.G单独使用效果并不佳。

关 键 词:D.S.G  多聚糖  烟草  TRIREAGENT  RNA提取

Application of New Reagent-D.S.G to Tobacco RNA Extraction and Purification
Li Zhengfeng,Li Wenzheng,Xia Yuzhen. Application of New Reagent-D.S.G to Tobacco RNA Extraction and Purification[J]. , 2007, 0(3)
Authors:Li Zhengfeng  Li Wenzheng  Xia Yuzhen
Affiliation:1 Yunnan Tobacco Research Institute,Yunnan Yuxi 653100; 2, Yunnan Agricultural University, Yunnan Kunming 650201; 3 Yunnan Yuxi Hongta Group Limited, Yunnan Yuxi 653100
Abstract:The extraction of nucleic acid is the foundation in molecular biological experiments, and RNA with high quality is the premise for cDNA library construction and genes expression. Five methods of RNA extraction, TRIREAGENT, D.S.G, guanidinium isothiocyanate and cesium chloride density gradient centrifugation, D.S.G and TRIREAGENT, were compared. The method of TRIREAGENT and D.S.G showed that applying the later two methods to extract tobacco RNA could wipe off the polysaccharide and other components of the tobacco cell wall. The result indicated that combination of D.S.G and TRIREAGENT was propitious extraction method of tobacco RNA with high quality. However, RNA quality wasn''t improved by D.S.G only.
Keywords:D.S.G   Polysaccharide   Tobacco   TRIREAGENT   RNA extraction
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