| 魔芋DNA快速微量提取及其ISSR-PCR扩增 |
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| 引用本文： | 杨朝柱,潘娅妮,谢玲玲,牟方贵,赵青华,马作江,冉瑞林,李明辉,滕建勋,陈永波,郭光耀,胡中立.魔芋DNA快速微量提取及其ISSR-PCR扩增[J].广西农业生物科学,2009(3):529-534. |
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| 作者姓名： | 杨朝柱 潘娅妮 谢玲玲 牟方贵 赵青华 马作江 冉瑞林 李明辉 滕建勋 陈永波 郭光耀 胡中立 |
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| 作者单位： | 恩施自治州农业科学院;武汉大学生命科学院;恩施市崔坝农技服务中心;湖北省农业厅油料作物中心; |
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| 基金项目： | 国家科技支撑计划项目(2007BAD73B03);;湖北省科技攻关项目(2007AA201C51)共同资助 |
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| 摘 要： | 魔芋属天南星科魔芋属草本植物，其组织富含多糖和多酚等次生物质，这类物质的存在不仅使DNA提取变得困难，还会影响下游的分子生物学操作。采用改进的提取方法，主要包括在2mL Eppendorf离心管中液氮研磨，CTAB-SDS裂解细胞，β-巯基乙醇浓度从2％提高到5％和添加PVP抑制多酚氧化，缓冲液去多糖等步骤，成功的提取了魔芋鲜叶的高质量DNA，其A260／A280位于1．80-2．00之间，产率超过470μg／g，ISSR0PCR扩增效果好。该方法具有快速、低廉、微量、稳定等特点，为深入研究魔芋资源遗传多样性、标记辅助育种和种芋纯度的鉴定奠定了基础。
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| 关 键 词： | 魔芋 组培苗 DNA提取 ISSR—PCR |
A Rapid DNA Minipreparation for Amorphorphallus Blume and Suitable for ISSR-PCR |
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| Yang Chaozhu , Pan Ya'ni Xie Lingling Mou Fanggui Zhao Qinghua Ma Zuojiang Ran Ruilin Li Minghui Teng Jianxun Chen Yongbo Guo Guangyao Hu Zhongli Agricultural Science Research Institution of Enshi Autonomic Prefecture,Enshi, College of Life Sciences,Wuhan University,Wuhan, Center of Oil Crop,Agricultural Ministry of Hubei Province, Cuiba Agrotechnical Service Center of Enshi City.A Rapid DNA Minipreparation for Amorphorphallus Blume and Suitable for ISSR-PCR[J].Journal of Guangxi Agricultural and Biological Science,2009(3):529-534. |
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| Authors: | Yang Chaozhu Pan Ya'ni Xie Lingling Mou Fanggui Zhao Qinghua Ma Zuojiang Ran Ruilin Li Minghui Teng Jianxun Chen Yongbo Guo Guangyao Hu Zhongli Agricultural Science Research Institution of Enshi Autonomic Prefecture Enshi College of Life Sciences Wuhan University Wuhan Center of Oil Crop Agricultural Ministry of Hubei Province Cuiba Agrotechnical Service Center of Enshi City |
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| Institution: | Yang Chaozhu 1,2 Pan Ya'ni 1 Xie Lingling 1 Mou Fanggui 1 Zhao Qinghua 1 Ma Zuojiang 1 Ran Ruilin 4 Li Minghui 3 Teng Jianxun 1 Chen Yongbo 1 Guo Guangyao 1 Hu Zhongli 2 1 Agricultural Science Research Institution of Enshi Autonomic Prefecture,Enshi,445000,2 College of Life Sciences,Wuhan University,Wuhan,430072,3 Center of Oil Crop,Agricultural Ministry of Hubei Province,430071,4 Cuiba Agrotechnical Service Center of Enshi City,445018 |
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| Abstract: | Amorphophallus blume, a herb plant belonging to the genus Amorphophallas in the family of araceae, contained plenty of polysaccharides, polyphenols and other undefined secondary metabolites, which resulted that DNA isolation became a challenge and the DNA polymers chains reaction(PCR) was failed, subsequently.A rapid, inexpensive and reliable DNA minipreparation method was developed and the DNA extracted from the fresh leaf of Amorphophallus blume was high quality and yield, and suitable for ISSR-PCR.The ra... |
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| Keywords: | Amorphophallus blume Tissue culture seedling DNA extraction ISSR-PCR |
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