| 单链DNA纯化对变性PAGE凝胶银染片段回收效率的提高 |
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| 引用本文： | 史金铭,孙野青.单链DNA纯化对变性PAGE凝胶银染片段回收效率的提高[J].生物学杂志,2009,26(5):74-76. |
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| 作者姓名： | 史金铭 孙野青 |
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| 作者单位： | 1. 哈尔滨工业大学生命科学与工程系,哈尔滨,150001
2. 哈尔滨工业大学生命科学与工程系,哈尔滨,150001;大连海事大学环境系统生物学研究所,大连,116026 |
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| 基金项目： | 国家高技术研究发展计划支持 |
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| 摘 要： | 为了提高变性聚丙烯酰胺凝胶电泳（PAGE）银染后片段回收的效率，本实验在传统的煮沸法的基础上加入单链DNA纯化的步骤对水稻基因组CCGG为点甲基化敏感限制性酶切多态性（MSAP）分析片段进行回收，并对加入该步骤后的再扩增的效率与传统煮沸法进行了比较。实验结果显示，加入单链DNA纯化步骤后的回收效率比传统煮沸法提高了约6倍。改进后的方法可以有效地用于扩增片段长度多态性（AFLP）以及MSAP等差异显示PAGE凝胶银染DNA片段的回收。
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| 关 键 词： | 变性聚丙烯酰胺凝胶电泳 条带回收 银染 MSAP |
ssDNA purification enhanced the fragments recovering from sliver stained denature PAGE gel |
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| SHI Jin-ming,SUN Ye-qing.ssDNA purification enhanced the fragments recovering from sliver stained denature PAGE gel[J].Journal of Biology,2009,26(5):74-76. |
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| Authors: | SHI Jin-ming SUN Ye-qing |
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| Institution: | SHI Jin-ming , SUN Ye-qing (1. Department of Life Science and Engineering, Harbin Institute of Technology, Harbin 150001; 2. Institute of Environmental Systems Biology, Dalian Maritime University, Dalian 116026, China) |
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| Abstract: | In order to enhance the efficiency of fragments recovering from sliver stained denature polyacrylamide gel electrophoresis （PAGE） gel, ssDNA purification was added to the traditional boiling method to recover the polymorphic fragments from MSAP analysis of methylation patterns on CCGG site of rice genome. The recovering efficiency of the improved method was measured with reamplification of PCR identification. Results showed that adding ssDNA purification enhanced the recovering efficiency about 6 times than the traditional boiling method. The improved method is more useful for recover the polymorphic fragments from PAGE gel of AFLP and MSAP analysis. |
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| Keywords: | MSAP |
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