| 钩距虾脊兰ISSR-PCR反应体系的建立与优化 |
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| 引用本文： | 彭德镇,孙小琴,孔令杰,李恩香,杨柏云.钩距虾脊兰ISSR-PCR反应体系的建立与优化[J].安徽农业科学,2009,37(31):15160-15162. |
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| 作者姓名： | 彭德镇 孙小琴 孔令杰 李恩香 杨柏云 |
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| 作者单位： | 南昌大学生命科学学院,江西南昌,330031;南昌大学生命科学学院,江西南昌,330031;南昌大学生命科学学院,江西南昌,330031;南昌大学生命科学学院,江西南昌,330031;南昌大学生命科学学院,江西南昌,330031 |
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| 摘 要： | 目的]为钩距虾脊兰种质资源研究奠定基础。方法]以钩距虾脊兰为试验材料，、利所改良的CTAB法提取钩距虾脊兰基因组DNA，并对影响ISSR扩增反应的各因素进行优化。结果]获得了高质量的钩距虾脊兰基因。组DNA；同时，建立了最适的钩距虾脊兰ISSR-PCR体系，即25山PCR反应体积中，2．5μl10×PCRbuffer，2．0mmol／LMgCl2，100ng模板DNA，240μmol／LdNTPs，1．75UTaqDNA聚合酶，0．4μmol／L引物；最佳扩增程序为94℃预变性5min。然后进行40个循环：94℃变性30s，复性温度根据各引物的TM值略低1～2℃，30s，72℃延伸50s，循环结束后72℃延伸7min，4℃保存。结论]这一优化系统的建立为进一步利用ISSR分子标记技术进行钩距虾脊兰遗传多样性研究提供了基础。
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| 关 键 词： | 钩距虾脊兰 DNA提取 ISSR分子标记 |
Establishment and Optimization of ISSR-PCR Reaction System for Calanthe graciliflora Hayata |
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| PENG De-zhen et al.Establishment and Optimization of ISSR-PCR Reaction System for Calanthe graciliflora Hayata[J].Journal of Anhui Agricultural Sciences,2009,37(31):15160-15162. |
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| Authors: | PENG De-zhen |
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| Institution: | PENG De-zhen et al(College of Life Sciences,Nanchang University,Nanchang,Jiangxi 330031) |
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| Abstract: | Objective]The research aimed to lay the foundation for the germplasm resources researches of Calanthe graciliflora Hayata.Method]With C.graciliflora as the test materials,the genomic DNA was extracted from C.graciliflora by using improved CTAB method.Different factors that affected ISSR amplification reaction were optimized.Result]High-quality genomic DNA was obtained from C.graciliflora Hayata.And the reaction system and amplified procedure that were suitable for C.graciliflora Hayata were as follows:25... |
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| Keywords: | Calanthe graciliflora Hayata DNA extraction ISSR |
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