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高质量植物基因组DNA的分离
引用本文:罗志勇,陈湘晖,等.高质量植物基因组DNA的分离[J].湖南医科大学学报,2001,26(2):178-180.
作者姓名:罗志勇  陈湘晖
摘    要:为了从富含多酚和我糖的药用植物组织中分离出高质量基因组DNA,本文通过改良几种已经存在的分离方法,获得了一种以CTAB法为基础的分离高质量完整DNA的简便、快速方法。使用该方法从人参、西洋参及三七的新鲜或干燥根组织分离DNA时,A260/A280为1.8左右,分子量大小约为21.2kb,由此所得的DNA可直接用于AFLP分析,即用EcoR I/Mse I消化DNA后,用DNA的限制酶切片段经T4 DNA连接酶连接,再用巢式PCR扩增,构建出可重复的、多态性丰富的人参、西洋参、三七基因组DNA AFLP指纹图谱。结果表明,该技术可有效地从富含多酚和多糖的药用植物组织中分离出高质量和高分子量DNA,此法亦有望用于其它植物DNA的分离。

关 键 词:DNA分离  高质量DNA  改进  CTAB法  药用植物

Isolation of high-quality genomic DNA from plants]
Z Y Luo,G Zhou,X H Chen.Isolation of high-quality genomic DNA from plants][J].Bulletin of Hunan Medical University,2001,26(2):178-180.
Authors:Z Y Luo  G Zhou  X H Chen
Institution:Molecular Biology Research Center, Xiangya Medical College, Central South University, Changsha 410078, China.
Abstract:In order to isolate high-quality genomic DNA from medicinal plant tissues enriching polyphenols and polysaccharides, a simple and rapid method based on CTAB extraction for isolating high-quality intact DNA was established by modifying several existing methods. With this technique, the absorbance ratio (A260/A280) of DNAs obtained from fresh and/or dried roots of Panax ginseng, P. Quinquefolius and P. notoginseng was 1.8 approximately. The restriction fragments of DNAs were directly digested with restriction enzyme (EcoR I/Mse I), linked up by T4 DNA ligase and amplified by nested PCR. Reproducible amplified fragment length polymorphism (AFLP) genomic DNA fingerprinting profiles were established with the isolated DNAs. The results demonstrate that the modified technique may be efficient and reliable in isolating high-quality and high-molecular-weight DNAs from fresh and/or dried medicinal plants containing a high content of polyphenols and polysaccharides. We expect that this method can also be applied to other plants.
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