假臭草DNA提取及RAPD反应体系的优化

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／LMg^2＋，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。

DNA Extraction and Optimization of RAPD Reaction System for Praxelis clematidea

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.