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《Acta Agriculturae Boreali-Sinica》 2008-03
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The Optimization of SRAP-PCR Amplification Program and System for Citrullus lanatus

LI Xiao-hui1,WANG Cong-yan2,XU Xiao-li1,CHANG Gao-zheng1,ZHANG Si-pu1(1.Institute of Horticulture,Henan Academy of Agriculture Sciences,Zhengzhou 450002,China;2.College of Life Sciences,Henan Agricultural University,Zhengzhou 450002,China)  
The SRAP-PCR reaction procedure and system for amplifying the watermelon genomic DNA was optimized using gradient experiment,so as to establish the optimum SRAP-PCR reaction conditions with high polymorphism,good repeatability and clear band pattern.The optimum SRAP-PCR reaction procedure was:pre-denaturation at 94℃ for 5 min followed by 5 cycles of denaturation at 94℃ for 1 min,anneal at 35℃ for 1 min and extension at 72℃ for 1 min;then 35 cycles of 94℃ for 1 min,50℃ for 1 min and 72℃ for 1 min;and a final extension at 72℃ for 5 min;kept at 4℃.The optimum SRAP-PCR system for a volume of 15 μL was:DNA 100 ng,Mg2+ 2.0 mmol/L,dNTPs 0.3 mmol/L,primer 60 ng,Taq polymerase 0.75 U.The procedure and system could meet the demands for genome SRAP amplification in C.lanatus.It was feable to apply the SRAP marker in genetic research in C.lanatus.
【Fund】: 河南自然科学基金(0611031800)
【CateGory Index】: S651
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