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《Journal of Fruit Science》 2009-02
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Establishment and optimization of ISSR reaction system for species of Eriobotrya

FU Yan1,LUO Nan1,YANG Qin1,2,WANG Yong-qing1,DENG Qun -xian1,LI Jun -qiang1,QIN Hong -mei1,RUAN Guang-lun3(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)  
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 ℃.
【Fund】: 四川省农作物育种攻关项目(项目编号:2001-08-03-13)
【CateGory Index】: S667.3
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