Comparison of Genomic DNA Extraction Methods for Chenopodium quinoa Willd  

藜麦基因组DNA提取方法的比较（英文）

作　　者：陆敏佳[1] 莫秀芳 王勤[1] 陆国权[1] 蒋玉蓉[1] 

机构地区：[1]浙江农林大学农业与食品科学学院,浙江杭州311300

出　　处：《Agricultural Science & Technology》2015年第7期1343-1347,1446,共6页农业科学与技术（英文版）

基　　金：Supported by National Natural Science Foundation of China(31301372);Key Project of Science and Technology Plan of Zhejiang Province(2011C12030);Innovation Training Project of Zhejiang Agriculture and Forestry University(201301004)~~

摘　　要：To rapidly obtain high-quality genomic DNA from Chenopodium quinoa Willd, the genomic DAN in different tissues （leaves, stems and roots） of Chenopodi- um quinoa Willd was extracted by modified CTAB method, SDS method and high- salt Iow-pH method, respectively. The quality and yield of extracted DNA was deter- mined using agarose gel electrophoresis and UV spectrophotometry. At the same time, the PCR-SSR and SSCP molecular detection was also performed. The results showed that the gel test strips, without obvious decomposition, of all the extraction methods were relatively obvious; the genomic DNA yield extracted by modified CTAB method was highest, followed by that by SDS method, and the genomic DNA extracted by high-salt Iow-pH method was lowest： the genomic DNA yields extracted by different methods from Chenopodium quinoa Wiltd leaves were all high- er than those from roots and stems; the quality of Chenopodium quinoa Willd ge- nomic DNA extracted by modified CTAB method and high-salt Iow-pH method was better, and polyphenols, polysaccharides and other impurities were removed more completely. The PCR-SSR and SSCP detection results showed that the genomic DNA extracted by different methods from different tissues of Chenopodium quinoa Willd all could be better amplified, and high-quality strips could be obtained. So the Chenopodium quinoa Willd genomic DNA extracted by the three methods all can be used for subsequent molecular biology research.为了快速获取高质量的藜麦基因组DNA,采用改良的CTAB法、SDS法和高盐低p H值法等3种方法分别提取藜麦不同组织(叶、茎、根部)的基因组DNA。通过琼脂糖凝胶电泳和紫外分光光度法测定比较所提DNA的质量和产量,同时进行了PCR-SSR、SSCP等分子检测。用不同方法提取藜麦不同组织部位DNA的结果表明:不同提取方法的凝胶检测条带均比较清晰,且无明显降解;改良的CTAB法所提取的DNA产率最高,SDS法其次,而高盐低p H值法最低;不同方法提取的叶片DNA产率均明显高于根部和茎部;改良的CTAB法和高盐低p H值法所提取的DNA质量较好,多酚类化合物和多糖等杂质去除得比较完全。PCR-SSR和SSCP检测结果表明:不同方法和不同组织所提取的DNA均能跑出良好的条带,适合进行后续的分子生物学研究。

关 键 词：Chenopodium quinoa Willd DNA extraction method： Molecular detection SSR： SSCP 

分 类 号：S519[农业科学—作物学]