摘要
建立白色念珠菌RAPD的最佳反应体系,并应用于其基因组DNA扩增。通过单因子试验分别研究了Mg2+、dNTPs、Taq酶、引物和模板DNA等浓度对RAPD反应的影响;同时,应用L16(45)正交试验研究了DNA模板、Mg2+、Taq酶、dNTPs和引物浓度对RAPD反应的影响。以条带稳定、丰富、清晰为标准,获得了白色念珠菌基因组DNA的RAPD扩增优化条件;对于白色念珠菌的最适RAPD反应体系为Mg2+1.5mmol/L、dNTPs 250μmol/L、引物0.6μmol/L、模板100ng/25μL、Taq DNA聚合酶1.5U/25μL。
To establish the optimization of RAPD reaction system for Candidia Albicans, the genomic DNA amplification was used for test. Effects of the concentrations of different substances like Mg^2＋ , dNTPs, DNA, primer, and Taq polymerase on RAPD amplification were studied through single factor experimentation. At the same time,the L16 （4^5） orthogonal experimentation was applied to study the effects of different concentrations of DNA, Mg^2 ＋ , Taq polymerase, dNTPs, and primer. Result showed the optimization of RAPD reaction system for Candidia Albicans was 1.5mmol/L Mg^2＋ , 250μ mol/L dNTPs, 0.6μ mol/L primer, 100ng/25μL template DNA, and 1.5U/25μL Taq DNA ploymerase.
出处
《生物学杂志》
CAS
CSCD
2010年第1期39-42,共4页
Journal of Biology
基金
皖南医学院中青年科研基金--不同基因型念珠菌感染临床株的分析
