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作 者:吴少慧[1] 刘忠[1] 张成刚[1] 张忠泽[1]
机构地区:[1]中国科学院沈阳应用生态研究所,沈阳110015
出 处:《应用与环境生物学报》2001年第1期76-78,共3页Chinese Journal of Applied and Environmental Biology
基 金:国家自然科学基金! (No :39870 0 38)资助项目
摘 要:建立了一种简捷、快速植物根瘤内痕量FrankiaDNA的提取方法 ,即采集自然态下根瘤 ,剥离瘤瓣 ,取瘤瓣尖经液氮研磨成粉末 ,以 2 0g/LCTAB(十六烷基三甲基溴化铵 )消化胞壁 ,然后以d =5~ 10 μm的石英粉吸附DNA ,再以无菌去离子蒸馏水释放DNA ,所获DNA产率及纯度较酚 /氯仿法好 .经对辽宁沙棘、色赤杨根瘤的瘤内痕量FrankiaDNA及体外培养的 9种Frankia菌株DNA分纯 ,并用于酶切及PCR ,获满意结果 .图 3参Actinomycete Frankia can form nitrogen-fixing symbiosis with a large number of woody dicotyledonous plants from different habits and demonstrates great diversity. A simple and rapid protocol was established and developed for purificating trace Frankia DNA from nodules. Frankia in nodules were digested with 20 g/L CTAB and released nucleic acid that were bound to silica particles (d=5~10 μm). Frankia DNA from 2 nodules and 9 cultured straing were purified with higher yields than those by phenol/chloroform, and could be used for restriction endonucleases and PCR. Fig 3, Ref 12
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