假臭草DNA提取及RAPD反应体系的优化  

DNA Extraction and Optimization of RAPD Reaction System for Praxelis clematidea

作　　者：李传军[1,2] 杨礼富[1] 王孝钢[3] 袁坤[1] 陈秋波[1] 易晓洁[1] 王真辉[1] 

机构地区：[1]中国热带农业科学院橡胶研究所/农业部橡胶树生物学重点开放实验室,海南儋州571737 [2]海南大学环境与植物保护学院,海南儋州571737 [3]农业部农产品质量安全监督检验测试中心(青岛),山东青岛266109

出　　处：《热带农业科学》2010年第5期21-25,共5页Chinese Journal of Tropical Agriculture

基　　金：国家自然科学基金项目(No.30960088)

摘　　要：以假臭草叶片为材料,对影响其随机扩增多态DNA(RAPD)反应的各因素进行优化,建立了假臭草RAPD的优化反应体系和程序,即在10μL反应体系中,5ng(/10μL)模板DNA,1.0μmol/L随机引物F15,150μmol/LdNTPs,2.0mmol/LMg2+,1.0UTaqDNA聚合酶;扩增程序为95℃预变性4min,95℃变性40s,36℃退火40s,72℃延伸1min,10个循环,后94℃变性30s,35℃退火30s,72℃延伸1min,35个循环,72℃延伸5min,4℃保温。The genomic DNA of Praxelis clematidea was employed to establish optimization of RAPD. The RAPD system for Praxelis clematidea was optimized including program, template, primer, dNTPs, Taq polymerase and Mg^2＋. Through ladder experiments, the reliable RAPD analysis system was established. The total reaction volume was 10 μL and reaction mixture consisted of template DNA, dNTPs, Mg^2＋, random primer and Taq DNA polymerase. RAPD program was 4 min at 95℃ for predenaturation, then 10 cycles of 40 s at 94℃ for denaturation, of 40 s at 36℃ for annealing, of 1 min at 72℃ for extension; then 35 cycles of 30 s at 94℃ for denaturation, of 30 s at 35℃ for annealing, of 1 min at 72℃ for extension, finally extension at 72℃ for 5 min.

关 键 词：假臭草 DNA提取 RAPD 体系优化 

分 类 号：S451[农业科学—植物保护]