Design and use of group-specific primers and probes for real-time quantitative PCR

作     者：Juntaek LIM Seung Gu SHIN Seungyong LEE Seokhwan HWANG 

作者机构：School of Environmental Science and EngineeringPohang University of Science and TechnologyPohangGyungbuk 790-784The Republic of Korea 

出 版 物：《Frontiers of Environmental Science & Engineering》 (环境科学与工程前沿（英文）)

年 卷 期：2011年第5卷第1期

页      面：28-39页

核心收录：

学科分类：0830[工学-环境科学与工程（可授工学、理学、农学学位）] 08[工学] 0812[工学-计算机科学与技术（可授工学、理学学位）] 

基　　金：This work was also supported by the Korea Research Foundation Grant funded by the Korean Government(MOEHRD) by New&Renewable Energy of the Korea Institute of Energy Technology Evaluation and Planning(KETEP)grant(No.20101T100100366) 

主　　题：absolute quantification design guideline primer probe real-time quantitative polymerase chain reaction(qPCR) 

摘      要：Real-time quantitative polymerase chain reaction(qPCR)has gained popularity as a technique to detect and quantify a specific group of target microorganisms from various environmental samples including soil,water,sediments,and *** qPCR is a very useful technique for nucleic acid quantification,accurately quantifying the target microbial group strongly depends on the quality of the primer and probe *** aspects of conducting qPCR assays have become increasingly routine and automated;however,one of the most important aspects,designing and selecting primer and probe sets,is often a somewhat arcane *** many cases,failed or non-specific amplification can be attributed to improperly designed primer-probe *** paper is intended to provide guidelines and general principles for designing group-specific primers and probes for qPCR *** demonstrate the effectiveness of these guidelines by reviewing the use of qPCR to study anaerobic processes and biologic nutrient removal *** assays using group-specific primers and probes designed with this method,have been used to successfully quantify 16S ribosomal Ribonucleic Acid(16S rRNA)gene copy numbers from target methanogenic and ammoniaoxidizing bacteria in various laboratory-and full-scale biologic *** with a good command of primer and probe design can use qPCR as a valuable tool to study biodiversity and to develop more efficient control strategies for biologic processes.