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《Journal of Southwest University(Natural Science Edition)》 2010-02
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Optimization of the ISSR-PCR System on Coreius heterodon(Bleeker) by Orthogonal Design

KONG Yan,ZHANG Hao-xing,XIE Xiao-junSchool of Life Science,Southwest University,Chongqing 400715,China  
The method of orthogonal design was used to optimize five factors,i.e.concentration of Mg2+,dNTP,primer,Taq DNA polymerase and DNA temple,which affect ISSR-PCR amplification results of Coreius heterodon(Bleeker),each at four levels,and an optimal ISSR-PCR reaction system for C.heterodon(Bleeker) was established: an ISSR-PCR reaction system with a total volume of 25 μL containing 1.5 mmol/L Mg2+,0.16 mmol/L dNTP,0.2 μmol/L primer,1U Taq DNA polymerase and 140 ng DNA temple.The testing results of the reaction system confirmed that it can produce stable and reproducible electrophoretic bands with high polymorphism of ISSR-PCR for C.heterodon(Bleeker).
【Fund】: 重庆市自然科学基金资助项目(CSTC2007BA7029)
【CateGory Index】: S917.4
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