| 三角紫叶酢浆草ISSR反应体系的建立与优化 |
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| 引用本文： | 胡甦,王永清,陶炼.三角紫叶酢浆草ISSR反应体系的建立与优化[J].草业学报,2011,20(5):142-150. |
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| 作者姓名： | 胡甦 王永清 陶炼 |
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| 作者单位： | 1.四川农业大学林学院,四川 雅安 625014;2.四川农业大学园艺学院,四川 雅安 625014;3.四川省农业科学院生物技术核技术研究所,四川 成都 610066 |
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| 基金项目： | 四川省育种攻关项目(2006YZGG-07-10)资助 |
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| 摘 要： | 采用正交设计直观分析法,对影响三角紫叶酢浆草ISSR-PCR反应较大的5个因素(Mg2+、Taq酶、引物、模板DNA、dNTP)在5个水平上进行优化实验;从100个随机引物中筛选出适宜的引物,并筛选出其最佳退火温度;最后对反应程序进行优化。由此首次建立了适合三角紫叶酢浆草的ISSR-PCR反应体系:25 μL反应液中含10×buffer 2.5 μL,Mg2+ 1.25 mmol/L,Taq酶0.9 U,引物0.3 μmol/L,模板DNA 60～100 ng,dNTP 0.25 mmol/L。反应程序为:94℃预变性5 min;94℃变性50 s,退火时间60 s(根据不同引物选择对应最佳退火温度),72℃延伸1.5 min,循环40次;72℃延伸7 min,4℃保存。这一优化系统的建立为今后利用ISSR标记技术研究酢浆草属植物提供理论依据和技术参考。
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| 关 键 词： | 三角紫叶酢浆草 ISSR 反应体系 |
Establishment and optimization of an ISSR reaction system for Oxalis triangularis |
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| HU Su,WANG Yong-qing,TAO Lian.Establishment and optimization of an ISSR reaction system for Oxalis triangularis[J].Acta Prataculturae Sinica,2011,20(5):142-150. |
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| Authors: | HU Su WANG Yong-qing TAO Lian |
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| Institution: | 1.College of Forestry, Sichuan Agricultural University, Ya’an 625014, China; 2.College of Horticulture, Sichuan Agricultural University, Ya’an 625014, China; 3.Institute of Biotechnology and Nuclear Technology, Sichuan Academy of Agricultural Sciences, Chengdu 610066, China |
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| Abstract: | An orthogonal design was used to optimize an ISSR(inter-simple sequence repeats)-PCR(polymerase chain reaction) amplification system for Oxalis triangularis,involving five levels of five main factors(Mg2+,Taq DNA polymerase,primer,template DNA and dNTP).The annealing temperature of suitable primers,which were selected from 100 random ISSR primers,was optimized and the reaction program was improved so that a suitable ISSR-PCR reaction system for O.triangularis was established.The reactions were performed in ... |
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| Keywords: | Oxalis triangularis ISSR reaction system |
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