实时定量PCR技术在本科实验教学中的应用

摘要

实时定量PCR技术是生命科学研究领域中的常用实验技术,为将其引入本科实验教学,课程组设计了"实时定量PCR分析不同浓度IPTG对启动子活性的调节作用"这一综合性实验内容.实验将RNA提取、反转录PCR、SYBR GreenⅠ染料法实时定量PCR和绝对定量分析串联起来,以获得启动子利用效率的变化情况,使学生通过实验教学能够深入、全面、系统地理解和掌握实时定量PCR技术,提高学生的综合能力和科研素养.%The real-time quantitative PCR(polymerase chain reaction)technology is the common experimental technology in the field of life science research.In order to introduce it into the undergraduate experimental teaching,the course group has designed the comprehensive experimental content,i.e.,"Analysis of regulation effect of IPTG different concentrations on promoter activity by real-time quantitative PCR ". In the experiment,the RNA extraction,reverse transcription PCR,SYBR Green I dye method,real-time quantitative PCR and absolute quantitative analysis are connected in series in order to obtain the change of the utilization efficiency of the promoter,which enables students to understand and grasp the real-time quantitative PCR technology and improve their comprehensive ability and scientific literacy through the experimental teaching.