佛手RAPD扩增系统的建立  

Establishment of RAPD amplification system in Bergamot

作　　者：马伯军[1] 陈镖[2] 陈文静[1] 张萍华[1] 

机构地区：[1]浙江师范大学化学与生命科学学院 [2]浙江大学城市学院

出　　处：《浙江师范大学学报（自然科学版）》2004年第2期158-161,共4页Journal of Zhejiang Normal University:Natural Sciences

基　　金：浙江省教育厅项目(20020850);浙江省大型仪器测试基金项目(03095)

摘　　要：试验了模板DNA、引物、镁离子和Taq酶的浓度对佛手RAPD分析结果的影响,从而建立了适合佛手RAPD扩增的系统.具体条件为:25μL反应体系中模板DNA100ng(4ng μL),MgCl23.0～5.0mmol L,dNTP200μmol L,10 merPrimer0.2μmol L及1UTaqPolymerase.扩增程序为:93℃120s;之后,93℃60s,35℃60s,72℃120s,进行42个扩增循环;最后72℃延伸600s.In this paper,the influencing factors on RAPD experiment were analyzed,such as the template concentration,the Mg^(2+)concentration,the primer concentration,the Taq polymerase concentration,and so on.The results indicated that each 25 μL amplification reaction was consisted of 100 ng template DNA,3～5 mmol/L Mg^(2+),200 μmol/L dNTP,0.2 μmol/L 10-mer primer,1 U Taq polymerase.The amplification procedure conditions were predenature at 93 ℃ 180 s followed by denature at (93 ℃) 60 s,annealing 35 ℃ for 60 s,extension 72 ℃ for 120 s,cycling for 42 times,last extension (300 s)and then the optimum procedure for Bergamot DNA amplification was obtained.

关 键 词：佛手 扩增 Taq酶 DNA 镁离子 引物 RAPD分析 反应体系 浓度 随机扩增多态DNA技术 

分 类 号：Q78[生物学—分子生物学] R282[医药卫生—中药学]