| 枇杷属植物ISSR反应体系的建立和优化 |
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| 引用本文： | 付燕,罗楠,杨芩,王永清,邓群仙,李俊强,秦红玫,阮光伦.枇杷属植物ISSR反应体系的建立和优化[J].果树学报,2009,26(2). |
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| 作者姓名： | 付燕 罗楠 杨芩 王永清 邓群仙 李俊强 秦红玫 阮光伦 |
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| 作者单位： | 付燕,罗楠,王永清,邓群仙,李俊强,秦红玫,FU Yan,LUO Nan,WANG Yong-qing,DENG Qun-xian,LI Jun-qiang,QIN Hong-mei(四川农业大学林学园艺学院,四川雅安,625014);杨芩,YANG Qin(四川农业大学林学园艺学院,四川雅安,625014;凯里学院环境与生命科学学院,贵州凯里,556000);阮光伦,RUAN Guang-lun(重庆市涪陵区果品办公室,重庆,408000)
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| 基金项目： | 四川省农作物育种攻关项目 |
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| 摘 要： | 首次通过正交实验,对影响枇杷属植物ISSR反应较大的Mg2+、Taq酶、dNTPs、引物、模板DNA浓度进行筛选,并对扩增反应程序进行优化。优化后的反应体系为:25μL反应体系中,含10×buffer2.5μL,Mg2+浓度2.0mmol·L-1,Taq酶1.5U,引物0.3μmol·L-1,模板DNA60ng,dNTPs0.15mmol·L-1。反应程序为94℃预变性5mim;94℃变性1mim,退火温度70s,72℃延伸1.5mim,40次循环;72℃延伸7mim,4℃保存。
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| 关 键 词： | 枇杷属植物 ISSR 正交设计 反应体系 |
Establishment and optimization of ISSR reaction system for species of Eriobotrya |
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| FU Yan,LUO Nan,YANG Qin,WANG Yong-qing,DENG Qun -xian,LI Jun -qiang,QIN Hong -mei,RUAN Guang-lun.Establishment and optimization of ISSR reaction system for species of Eriobotrya[J].Journal of Fruit Science,2009,26(2). |
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| Authors: | FU Yan LUO Nan YANG Qin WANG Yong-qing DENG Qun -xian LI Jun -qiang QIN Hong -mei RUAN Guang-lun |
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| Institution: | 1College of Forestry and Horticulture;Sichuan Agricultural University;Ya'an;Sichuan 625014 China;2College of Environmental and Life Science;Kaili University;Kaili;Guizhou 55600 China;3Fruit Office of Fuling Area of Chongqing;Chongqing 408000 China |
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| Abstract: | The concentration of Mg2+,Taq polymerase,dNTPs,primer and template DNA,greatly influencing ISSR-PCR of loquat,were optimized by orthogonal design in this study,and the reaction procedure was also improved.Thus an optimized ISSR reaction system was established for the first time for loquat.The total 25 μL contained 10×buffer 2.5 μL,2.0 mmol·L-1 Mg2+,1.5 U Taq polymerase,0.3 μmol·L-1 primer,60 ng template DNA,and 0.15 mmol·L-1 dNTPs.The reaction procedure was:94 ℃ for 5 min,followed by 40 cycles of 94 ℃ for 1 min,annealing temperature for 70 s,72 ℃ for 1.5 mim,and was ter-minated with a 7 min extension step at 72 ℃. |
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| Keywords: | ISSR |
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