摘要
以焦旱1号总DNA为材料,首先对影响RAPD-PCR反应的模板DNA、Mg2+、dNTP、引物和TaqDNA聚合酶浓度等因素进行了初步优化,分析了各因素对RAPD-PCR扩增结果的影响。在此基础上对影响RAPD-PCR反应的Mg2+、dNTP、引物和TaqDNA聚合酶浓度等4个主要因素进行正交优化,研究结果表明:在25μlRAPD-PCR反应体系中,模板DNA20ng;Mg2+浓度1.5mmol/L;dNTP的浓度0.2mmol/L;引物量15pmol;TaqDNA聚合酶1.0U。在此最佳条件下,利用引物B8对18个北方粳稻品种进行了成功的扩增。
It was to obtain the optimum RAPD-PCR reaction system for Jiaohan No. 1 ,one test was designed to screen the suitable concentration of 4 factors （ Mg^2＋ , dNTP, primer, Taq）in the reaction system. The optimal protocol was accomplished by orthogonal method in 25μl reaction volumes containing 20 ng template DNA, 1.5 mmol/L MgCl2 ,0. 2 mmol/L dNTPs, 15 pmol each primer and 1.0 unit Tag polymerase. The results showed that primers Bs can be used to amplify 18 species of Japonica Rice.
出处
《生物技术通报》
CAS
CSCD
北大核心
2009年第12期92-95,101,共5页
Biotechnology Bulletin
