蓝萼香茶菜随机扩增多态DNA(RAPD)反应体系优化  

Optimization of RAPD System of Isodon japonica (Burm.f.) Hara var.glaucocalyx (Maxim.) Hara

作　　者：金忠民[1] 沙伟[1] 孙天国[1] 孙雪巍[1] 

机构地区：[1]齐齐哈尔大学生命科学与工程学院,黑龙江齐齐哈尔161006

出　　处：《广西科学》2004年第3期258-260,共3页Guangxi Sciences

基　　金：齐齐哈尔市科技局计划项目

摘　　要：试验以香茶菜属植物的蓝萼香茶菜 [Isodon japonica (Burm.f.) Hara var.glaucocalyx (Maxim.) ]为材料 ,采用改进的 CTAB法提取基因组 DNA,首次建立适合于蓝萼香茶菜的 RAPD(Random amplifiedpolymorphic NDA,随机扩增的多态性 )分析的 PCR反应体系。通过对 RAPD条件优化分析 ,确定蓝萼香茶菜RAPD的最佳方案为 :2 5μl反应体系中包括三磷酸脱氧核苷酸 (d NTPs) 0 .1mm ol/ L ,引物 0 .4μmol/ L ,模板DNA90 ng,Taq酶 (Sangon) 3u,2 .5 μl10×buffer缓冲液 (含 2 0 m mol/ L Mg2 + ) ,其余部分用无菌超纯水补平 ;PCR扩增循环为 93℃预变性 4 min,93℃变性 4 5 s,36℃复性 75 s,72℃延伸 2 min,4 0个循环 ,最后 72℃延伸 10 min。The total DNA from the young leaves of I.japonica (Burm.f.) Hara var.glaucocalyx (Maxim.) Hara was successfully extracted by using the developed CTAB method and a steady system of PCR reaction for I.japonica(Burm.f.) Hara var.glaucocalyx (Maxim.) Hara was built.The optimal PCR system for RAPD analysis was as follows:0.1 mmol/L dNTPs,0.4 μmol/L random primer,90ng DNA template,3u Taq polymerase in 25 μl reaction solution.The reaction program was devised for one cycle at 93℃ 4 minutes and followed by 40 cycles, each with 45 seconds at 93℃,75 seconds at 37℃,2 minutes at 72℃,and a final extension at 72℃ for 10 minutes.

关 键 词：蓝萼香茶菜 RAPD 优化 

分 类 号：Q943[生物学—植物学]