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Optimization of SSR-PCR system of spaceflight alfalfa and primers selection

FAN Run-jun1,CHON Ben-jian1,DENG Bo2(1.College of Pratacultural Science,Gansu Agricultural University,Lanzhou 730070,China;2.Dept.of Grassland Science,College of Animal Science and Technology,China Agricultural University,Beijing 100193,China;3.Gansu Tianshui Agricultural Sciences Research Institute,Tianshui,741000,China)  
Spaceflight breeding has high variation frequency,large variation amplitude,more profitable variation and high stability.The orthogonal design was used to optimize SSR amplification system of spaceflight alfalfa.Two orthogonal experiments were conducted to investigate the amplification efficiency under different amplification conditions.Results showed that the optimum concentrations of reactants in 25 μl reaction mixture were as follows:dNTP0.2 mmol/L,Taq DNA polymerase 2U,Primers 0.7 μmol/L,Mg2+2.5 mmol/L,10×buffer and 60 ng template DNA.Totally 6 SSR primers were selected from 17 SSR primers by sieving which could produce DNA polymorphism.
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