【作者】 陈惠云； 孙志栋； 茅轶俊； 王晓武； 葛红；

【Author】 Chen Hui-Yun, Sun Zhi-Dong, Mao Yi-Jun, Wang Xiao-Wu, Ge Hong) (Ningbo Academy of Agricultural Sciences, Ningbo, China. 315040. China Academy of Agricultural Sciences, vegetable and flower institute, Beijing, china. 100081)

【机构】 宁波市农科院； 中国农科院蔬菜花卉研究所；

【摘要】 本文通过比较不同的DNA提取方法、春兰材料不同部位的基因组DNA提取质量,试图找出春兰(Cymbidium goeringii(Rchb.f.)Rchb.f.)基因组DNA提取的最佳方法,为更好开展春兰分子生物学研究奠定技术基础。结果发现,CTAB法的两种化学药品,即十六烷基三甲基溴化铵(Cetyl-trimethyl-Ammonium Bromide,上海产)与十六烷基二甲基乙基溴化铵(Cetyldimethylethyl -ammonium bromide,美国BBI产)之间提取得率和质量差异不显著。而SDS法的两种不同缓冲液配制方法对春兰基因组DNA提取有显著差异。比较CTAB法和SDS法,改良的SDS法提取的春兰DNA质量与得率比改良的CTAB法相对要好,但就花药而言,还是改良的CTAB法(采用进口CTAB)较好,去除蛋白质相对干净。春兰材料的不同提取部位也有差异,花苞基因组DNA提取得率和质量优于叶片。此外,实验中发现,适当增加氯仿/ 异戊醇抽提次数、延长异丙醇的沉淀析出时间、水浴时间等可以提高春兰基因组DNA的提取质量。更多还原

【Abstract】 In our study, we try to find out the optimal method of extracting the genome DNA from Cymbidium goeringii (Rchb. f.) Rchb. f. so as to lay foundation of the molecular biology through comparing the extraction quality of different extraction methods and different parts within the Cymbidium. The result showed that there was no significant difference between two kinds of chemical medicines for CTAB(Cetyl-trimethyl-Ammonium Bromide from Shanghai in China and Cetyl-dimethylethyl-ammonium bromide from BBI in USA.),and significant difference between two kinds of buffers for SDS. And the improved method for SDS was better than the one for CTAB in the quality of the genome DNA extracted from Cymbidium. But speaking of the flower, improved CTAB methods (imports CTAB) was better than other methods because of removing the protein relatively clean. In addition, it was founded that the quality of genome DNA would be improved by increasing extraction times for the Isoamyl-Chloroform, extending deposition time for lsopropylalcohol, and extending water bath time.更多还原