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《Plant Protection》 2014-02
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Establishment and optimization of ISSR-PCR reaction system with orthogonal design for Sogatella furcifera

Xie Jianan;Guo Jianjun;Jin Daochao;Provincial Key Laboratory for Agricultural Pest Management of Mountainous Regions,Institute of Entomology,Guizhou University;  
Sogatella furciferais one of the most important rice pests.To establish and optimize ISSR-PCR reaction system for S.furcifera,the factors influencing ISSR system were studied with L16(45)orthogonal design,and the optimal annealing temperature and cycle times were proposed.The results showed that the optimal conditions for ISSR-PCR system were 2μL10×PCR buffer(10 mmol/L Tris-HCl,50 mmol/L KCl),1.5 mmol/L Mg2+,0.25 mmol/L dNTPs mixture,0.9μmol/L of each primer,1U Taq DNA polymerase(TaKaRa)and 50 ng template DNA in 20μL reaction volume.The optimal annealing temperatures were 51.0-54.0℃for the 10 primers used in the present study.The clear and reproducible DNA bands were obtained by the optimal annealing temperature and 40 reaction cycles.
【Fund】: 贵州省农业攻关(黔科合NY字[2010]3064号);; “973”计划前期研究专项(2009CB125909);; 贵州省教育厅自然科学研究项目(2010011)
【CateGory Index】: S435.112.3
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