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《Acta Agriculturae Boreali-Sinica》 2008-S2
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Optimization of SRAP Reaction System in Peach(Prunus persica)

SHI Hong-li,HAN Ming-yu,ZHAO Cai-ping(College of Horticulture,Northwest Agricultural and Forestry University,Yangling 712100,China)  
The aim of the research was to establish SRAP-PCR amplification system which was suitable for Peach(Prunus persica) genome DNA so as to lay foundation for the construction of gene map and molecular marker in Peach(Prunus persica).Peach(Prunus persica) genome DNA as template,the major components of SRAP,such as concentrations of dNTPs,Mg2+,Taq DNA polymerase,primers and template,were optimized in this study by orthogonal design in five factors four levels respectively.The results showed that the optimum SRAP reaction system includes dNTPs 0.12 mmol/L,Mg2+ 4 mmol/L,Taq DNA polymerase 2 U,primer 0.3 mmol/L and DNA template 50 ng in the 25 μL system.The most suitable protocol was initially denaturing at 94℃ for 5 min,then pre-amplifying at 94℃ 1 min,35℃ l min and 72℃ 1 min for five cycles,finally amplifying for 35 cycles when the annealing temperature was adjusted to 50℃.
【Fund】: 陕西省攻关项目(2006K01-G27-01);; 农业部“948”项目(2006-G27)
【CateGory Index】: S662.1
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