摘要
目的:对RAPD的反应体系进行优化,以期建立稳定的临床检测系统,在控制院内感染的流行病研究中发挥更大的作用。方法:用RAPD技术进行基因分型。结果:引物浓度:浓度为0·5μmol/L时扩增最好。DNA模板:浓度在0·50ng/μl、2ng/μl均得到了良好的扩增效果。镁离子浓度:在2·0mmol/L、4mmol/L时扩增效果较好。退火温度:在30℃时扩增条带最清晰。循环参数:在35、45个循环时扩增效果均好。结论:本试验对RAPD反应条件进行优化,建立了较稳定的临床检测体系。
Objective： to optimize the Randomly Amplified DNA Polymorphism （RAPD） reaction system, and set up a stable clinical monitoring system （for） cross infection. Methods：Using RAPD technique for gene classifing. Results： optimization results by RAPD reaction system：When the density of the primer was 0. 5μmol /L, amplification would be the best; when the density of DNA template was 0.5ng/μl or 2ng/μl, the amplification was also better; when magnesium ion density was 2.0mmol/L or 4.0 mmol/L, the effect of amplification was comparatively better. Temperatures of anneal： At 30℃, the stripes of amplification were most distinctive. Parametre＇s of cycles： Good amplification would be achieved at both 35 and 45 cycles. Conclusion： In this experiment, by optimizing the RADP reaction system, a stable clinical examination system can be obtained, which played an important role in the investigation of hospital infection epidemiology.
出处
《河北北方学院学报（医学版）》
2005年第4期1-3,共3页
Journal of Hebei North University:Medical Edition
基金
河北省自然科学基金指导项目(编号:012761169)
关键词
随机扩增DNA多态性分析
反应体系优化
细菌
Randomly Amplified DNA Polymorphism Analyse
Optimization Reaction System, Bacterium
