实时荧光定量PCR技术在实验教学中的应用  

Application of real-time fluorescent quantitative PCR technology in experimental teaching

作　　者：赵玉红[1] 李欣[1] 赵立青[1] 李小菊[1] 石建党[1] 李登文[1] 张金红[1] 周浩[1] Zhao Yuhong;Li Xin;Zhao Liqing;Li Xiaoju;Shi Jiandang;Li Dengwen;Zhang Jinhong;Zhou Hao(Biological Experimental Teaching Center,Nankai University,Tianjin 300071,China)

机构地区：[1]南开大学生物实验教学中心,天津300071

出　　处：《实验技术与管理》2018年第4期61-64,68,共5页Experimental Technology and Management

基　　金：国家基础学科人才培养基金"条件建设项目"(J1310003);南开大学生命科学学院2017年教学改革项目

摘　　要：设计了"实时荧光定量PCR分析IPTG诱导EGFP基因在大肠杆菌中的异源表达"实验,研究IPTG诱导浓度和温度对EGFP基因表达的影响。结果表明,IPTG浓度为270μmol/L、诱导温度为16℃、诱导时间为18h的条件下,EGFP mRNA的表达量最大。通过该实验的学习,使学生掌握实时荧光定量PCR的相关实验原理、技术以及实时荧光定量PCR仪的使用,培养学生的科研能力和综合素质。The experiment on the real-time fluorescence quantitative PCR(polymerase chain reaction)analysis of the heterologous expression of EGFP gene induced by IPTG in Escherichia coli,is designed,and the effect of IPTG induction concentration and temperature on the expression of EGFP gene is studied.The preliminary results show that under the conditions that the concentration of IPTG is 270μmol/L,the induction temperature is 16℃and the induction time is 18 h,the expression amount of EGFP mRNA is maximal.Through the study of this experiment,students can master the related experimental principles and techniques of real-time fluorescent quantitative PCR and the use of real-time fluorescent quantitative PCR.The students’scientific research ability and comprehensive quality are trained.

关 键 词：实时荧光定量PCR 核酸定量 IPTG 

分 类 号：Q5-33[生物学—生物化学] Q503