大百合RAPD体系的建立与优化  

Establishment and optimization of Cardiocrinum giganteum's RAPD system

作　　者：喻晓[1] 文涛[1] 曾杨[2] 

机构地区：[1]四川农业大学农学院,四川雅安625014 [2]四川农业大学林学园艺学院,四川雅安625014

出　　处：《西南农业学报》2007年第6期1304-1306,共3页Southwest China Journal of Agricultural Sciences

摘　　要：以大百合DNA为模板,采用正交试验设计,对影响大百合RAPD-PCR扩增的重要参数进行了优化试验,以期建立大百合RAPD反应的优化体系。结果表明最优大百合RAPD-PCR的反应体系(20μl):Mg2+(2.0 mmol/L)1.0μl、dNTPs(0.2 mmol/L)1.0μl、primer(0.6 umol/L)1.3μl、DNA(30 ng/μl)1.0μl、Taq酶1U、10×buffer2μl;扩增程序为:在94℃下预变性5 min,然后进行35个循环(94℃变性30s、37℃退火50s、72℃延伸1 min),最后在72℃延伸10 min,在电压为50 V下电泳1 h。To use the DNA of Cardiocrinum giganteum as a template to do the optimization test on the important parameters of the RAPD-PCR amplification for Cardiocrinum giganteum the Cardiocrinum giganteum＇ s RAPD optimization system, and the use of orthogonal design method, the Cardiocrinum giganteum＇ s RAPD-PCR conditions were optimized. The results showed that the best RAPD-PCR reaction system （20 g,1 ） must follow those conditions as bellows： Mg^2＋ ＋ （ 2.0 mmoVL ） 1.0 μl, dNTPs （ 0.2 mmol/L） 1.0 μ1, primer （ 0.6 μmol/l ） 1.3 μl, DNA （ 30 ng/μl） 1.0 μl, Tag enzyme 1 U, 10 × buffer 2 μl. PCR procedure was discribed as bellow： to be predegenerated at 94 ℃ for 5 rain, then be followed by 35 cycles（ denaturation at 94℃ for 30 s,annealing at 37 ℃ for 50 s,extensfon at 72℃ for 1 min） ,finally to be extended at 72 ℃ for 10 min and electro-phoresis one hour in 50V.

关 键 词：大百合 RAPD 遗传多样性 milli-Q 

分 类 号：S664.3[农业科学—果树学]