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《Journal of Nuclear Agricultural Sciences》 2007-05
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ESTABLISHMENT OF AN ISSR REACTION SYSTEM FOR POPLAR AND OPTIMIZATION USING ORTHOGONAL DESIGN

WANG Jie-ming XIANG Yan WU Da-qiang SUN Zhi-juan CAI Cheng(College of Forest and Garden,Anhui Agricultural University,Hefei,Anhui 230036)  
Orthogonal design was applied for optimizing five factors(Taq DNA polymerase,Mg~(2+),DNA template,dNTP,and primer) in the ISSR-PCR amplification system on poplar respectively.As a result,a satisfactory ISSR reaction system for poplar with desirable repeatability and polymorphic bands was established.In a total volume of 25μl ISSR-PCR system,it contained 1×buffer,0.12mmol/L dNTP,0.4μmol/L primer,3.5 mmol/L Mg~(2+),1.5U Taq DNA polymerase and 40ng template DNA.The optimal annealing temperature of every prime for ISSR-PCR reaction was proposed by gradient PCR.It was found that different primers had different annealing temperature.As far as concerned with primer W95142,the optimal annealing temperature was 56.1℃.This optimized ISSR reaction system would provide the basis for the analysis of diversity,map construction and gene localization of important traits of poplar with ISSR markers.
【Fund】: 安徽省科技厅自然基金(03041109)
【CateGory Index】: S792.11
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