摘要
白背飞虱Sogatella furcifera(Horváth)是水稻重要害虫之一。研究采用L16(45)正交设计优化试验,构建了白背飞虱RAPD最优反应体系,并对引物退火温度和反应循环次数进行了优化。结果表明:白背飞虱RAPDPCR最优反应体系为2μL 10×PCR Buffer(10 mmol/L Tris-HCl;50 mmol/L KCl),2.4 mmol/L Mg2+,0.15 mmol/L dNTPs,0.7μmol/L引物,0.8U Taq DNA Polymerase和30 ng模板DNA,ddH2O补充至20μL。本研究采用的10条引物的最佳退火温度在35-39℃间,以引物相应最优退火温度和40次PCR反应循环可获得较好的白背飞虱RAPD-PCR结果。
Sogatellafurcifera is one of the most important pests of rice. To establish and optimize RAPD - PCR reaction system for S. furcifera, the factors influencing RAPD 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 RAPD - PCR system consists of 2 μL 10 × PCR buffer ( 10 mmol/L Tris- HC1; 50 mmol/L KC1), 2.4 mmol/L Mg2+, 0. 15 mmol/L dNTPs mixture, 0.7μmol/L primer, 0. 8U Taq DNA polymerase and 30 ng DNA template, and adds ddH20 to 20 μL in total. The optimal annealing temperature is 35 -39 ℃ for 10 primers used in the present study. The clear and reproducible DNA bands were obtained by the optimal annealing temperature and the 40 reaction cycles.
出处
《环境昆虫学报》
CSCD
北大核心
2014年第3期348-353,共6页
Journal of Environmental Entomology
基金
973计划前期研究专项(2009CB125908)
贵州省农业攻关(黔科合NY字[2010]3064号)
贵州省教育厅自然科学研究项目(2010011)
关键词
白背飞虱
正交设计
RAPD
Sogatella furcifera
orthogonal design
RAPD