| 基于棉花BAC文库的大片段核DNA的提取方法 |
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| 引用本文： | 殷剑美,张天真.基于棉花BAC文库的大片段核DNA的提取方法[J].分子植物育种,2004,2(3):354-357. |
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| 作者姓名： | 殷剑美 张天真 |
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| 作者单位： | 南京农业大学作物遗传与种质创新国家重点实验室,南京,210095;南京农业大学作物遗传与种质创新国家重点实验室,南京,210095 |
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| 摘 要： | 本实验在基因组大片段DNA的提取方法上，结合棉花的特殊性，作了一些改进。即首先将幼苗进行暗培养，减少了叶绿体和酚类等物质的影响；随后在Wash buffer中加入酚结合剂PVP40(polyvinylpyrrolidone)，较好地去除了棉酚等次生物质的干扰；另外在细胞核分离过程中，增加了低速稍离心去沉淀的步骤但之前不需要过滤，较好地去除了不溶性杂质。此方法所提取得到的棉花基因组DNA，分子量约在1Mb左右，碎片较少，基本上无蛋白质和细胞器DNA等污染。其质量完全满足构建BAC文库的要求。
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| 关 键 词： | 大片段DNA 提取 BAC文库 棉花 |
Preparation of Meg-base DNA Cotton Nuclei for BAC Library |
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| Yin Jianmei Zhang Tianzhen
National Key Laboratory of Crop Genetics and Germplasm Enhancement,Nanjing Agricultural University,Nanjing.Preparation of Meg-base DNA Cotton Nuclei for BAC Library[J].Molecular Plant Breeding,2004,2(3):354-357. |
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| Authors: | Yin Jianmei Zhang Tianzhen
National Key Laboratory of Crop Genetics and Germplasm Enhancement Nanjing Agricultural University Nanjing |
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| Institution: | Yin Jianmei Zhang Tianzhen
National Key Laboratory of Crop Genetics and Germplasm Enhancement,Nanjing Agricultural University,Nanjing,210095 |
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| Abstract: | Some methods are used to improve the preparation meg-base DNA cotton nuclei for BAC library. The seedlings grew under lightless to reduce effects of chloroplast and polyphenol etc.; PVP40 was added to excluded polyphenol and other secondary interfacing substance; additionally, to remove the particulate matter remaining in the suspension, filter the re-suspended nuclei into a centrifuge tube through two layers of Miracloth and centrifuge the contents at low speed and short time during the preparation of intact nuclei. It showed that the resulting DNA, about 1Mb and almost free protein and organelle DNA, is suitable for the construction of cotton BAC library. |
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| Keywords: | Meg-base DNA Preparation BAC library Cotton |
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