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《Anhui Agricultural Science Bulletin》 2007-09
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A Studies on DNA Extraction and Establishmen of RAPD Reaction System in Achnatherum splendens

Xu Haixia,Zhang Xia,Yan Ping,Du Jinzhou(College of Biological Engineering,Shihezi University,Shihezi,Xinjiang 832003,China)  
Genomic DNA of in A.splendens was successfully extracted by CTAB.Factors influencing RAPD result,including the concentration of DNA template,Mg2+,dNTP,primers and Taq,were studied.An optimul PCR system for RAPD analysis was found: in 12.5μL reaction solution,contained 10mmol·L-1 Tris-HCl(pH8.0),50mmol·L-1KCl,1.50mmol·L-1MgCl2,0.2mmol·L-1 dNTP,3.2pmol random primer,20ng DNA template,0.5U Taq polymerase.The amplification program was devised: denaturing at 95℃ for 30s,primer annealing at 35℃ for 45s,extension at 72℃ for 80s,6 cycles;then 94℃ 45s,36℃ 40s,72℃ 70s,36 cycles;and 10min at 72℃ in the final extension.The reaction system was well reproducible and highly rebiable,and it couble be effectively for RAPD analysis in the systematic study in A.splendens.
【Fund】: 国家重大基础研究前期研究专项资助项目(2004CCA02800);; 新疆兵团绿洲生态农业重点实验室资助项目(200313);; 石河子大学科学研究青年基金项目(200257)
【CateGory Index】: S543.9;S561
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