| 条斑紫菜高纯度总DNA及其质粒状DNA的提取 |
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| 引用本文： | 郭宝太,毕玉平,单雷,李广存,戴继勋.条斑紫菜高纯度总DNA及其质粒状DNA的提取[J].海洋学报,2000,22(2):87-91. |
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| 作者姓名： | 郭宝太 毕玉平 单雷 李广存 戴继勋 |
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| 作者单位： | 1.青岛海洋大学海洋生命学院, 青岛266003 |
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| 摘 要： | 提取条斑紫菜高纯度总DNA及其质粒状DNA的新方法。先用海螺酶处理紫菜叶状体制备细胞,然后用SDS-蛋白酶K裂解细胞提取总DNA,再用玻璃粉浆(glassmilk)对其纯化,经纯化后的总DNA能被EcoRI,Dral与HaeⅢ等限制酶完全酶切,并在酶切图谱上形成明显的DNA带型。当用异硫氰酸胍一十二烷基肌氨酸钠裂解紫菜细胞时,在总DNA提取物中直接发现有一条质粒状DNA带(2.3Kb),即建立了一种极简便的质粒状DNA提取方法。
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| 关 键 词： | 条斑紫菜 细胞 总DNA glassmilk 完全酶切 质粒状DNA |
| 收稿时间： | 1998/12/26 0:00:00 |
| 修稿时间： | 1998-12-26 |
Extraction of plasmid-like DNA and high-quality total DNA from Porphyra yezoensis |
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| Guo Baotai,Bi Yuping,Shan Let,Li Guangcun and Dai Jixun.Extraction of plasmid-like DNA and high-quality total DNA from Porphyra yezoensis[J].Acta Oceanologica Sinica (in Chinese),2000,22(2):87-91. |
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| Authors: | Guo Baotai Bi Yuping Shan Let Li Guangcun and Dai Jixun |
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| Institution: | 1.College of Marine Life Science, Ocean University of Qingdao, Qingdao 2660032.Biotechnology Research Center, Shandong Academy of Agricultural Science, Jinan 250100 |
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| Abstract: | Cellls were prepared from sea snail enzyme digests of Porphyra yezoensis Thalli. Crude extracts of total DNA were isolated from Porphyra cells lysed with SDS-Proteinase K solution. Total DNA was purified with glassmilk, the resulting DNA was of high quality, could be completely digested with restriction enzymes. So that it was also suitable for analyses of Southern hybridization, RFLP and AFLP, etc. DNA bands were clearly observed in the agarose gel electrophoresis patterns of total DNAs digested with EcoRI, PstI and HaeⅢ respectively. The presence of DNA bands in the electrophoresis patterns of total DNA showed that the nuclear genome of Porphyra yezoensis may be small. Unexpectedly, a plasmid-like DNA band (2.3Kb) was found in the alectrophoresis pattern of P. Yezoensis total DNA isolated with guanidinium thiocyanate-sarcosyl solution. This is a very simple and convenient method for plasmid-like DNA isolation. |
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| Keywords: | Porphyra yezoensis cells total DNA glassmilk complete digestion plasmid-like DNA |
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