| 从水稻细胞核中制备分子量达到Mb级的DNA |
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| 引用本文： | 王春新,谢亦武,刘良式. 从水稻细胞核中制备分子量达到Mb级的DNA[J]. 生物化学与生物物理进展, 1995, 22(4): 357-361 |
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| 作者姓名： | 王春新 谢亦武 刘良式 |
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| 作者单位： | 广州中山大学生命科学学院;广州中山大学生命科学学院;广州中山大学生命科学学院 |
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| 基金项目： | 国家“863”计划(863-101-01-03)及美国洛克菲勒基金会 (1994-0001-0137)资助. |
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| 摘 要： | 用水稻黄化苗作实验材料,经蔗糖不连续梯度纯化细胞核、琼脂糖包埋、蛋白酶消化释放DNA,脉冲交变电泳显示所得DNA样品分子量在200kb至3Mb之间,其中大量集中在2.2Mb处.所得DNA容易被多种内切酶消化和连接,可用于构建水稻YAC文库和大尺度物理图的研究.
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| 关 键 词： | 水稻 Mb级DNA 脉冲交变电泳 |
| 收稿时间： | 1994-07-26 |
| 修稿时间： | 1994-12-19 |
Isolation of Megabase DNA from Rice Nuclei |
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| Wang Chunxin,Xie Yiwu and Liu Liangshi. Isolation of Megabase DNA from Rice Nuclei[J]. Progress In Biochemistry and Biophysics, 1995, 22(4): 357-361 |
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| Authors: | Wang Chunxin Xie Yiwu Liu Liangshi |
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| Affiliation: | School of Life Science, Zhongshan University;School of Life Science, Zhongshan University;School of Life Science, Zhongshan University |
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| Abstract: | A convenient and efficient procedure for isolation of rice HMW DNA bad been developed.Etiolated seedings were ground into the fine powder with a pestle and mortar under liquid nitrogen,then extracted with a protective medium,Filtrate was layered on discontinuous sucrose gradient,then the purified nuclei were embeded in agarose gel.After proteinase digestion,the HMW DNA was released. Pulsed-field gel electrophoresis showed that the sample size ranges from 200 kb to 3 Mb and in the majority of 2.2 Mb. Rice YAC library was constructed successfully, using these easily digestable DNA preparation. |
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| Keywords: | rice Mb DNA pulsed field ge1electrophoresis(PFGE) |
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