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Juntaek LIM Seung Gu SHIN Seungyong LEE Seokhwan HWANG 《Frontiers of Environmental Science & Engineering》2011,5(1):28-39
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 sludge. Although 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 used. Many 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 process. In many cases, failed or non-specific amplification can be attributed to improperly designed primer-probe sets. This paper is intended to provide guidelines and general principles for designing group-specific primers and probes for qPCR assays. We demonstrate the effectiveness of these guidelines by reviewing the use of qPCR to study anaerobic processes and biologic nutrient removal processes. qPCR 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 ammonia-oxidizing bacteria in various laboratory- and full-scale biologic processes. Researchers 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. 相似文献
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Juntaek Lim Seung Gu Shin Seungyong Lee Seokhwan Hwang 《Frontiers of Environmental Science & Engineering in China》2011,5(1):28-39
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 sludge. Although 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 used. Many 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 process.
In many cases, failed or non-specific amplification can be attributed to improperly designed primer-probe sets. This paper
is intended to provide guidelines and general principles for designing group-specific primers and probes for qPCR assays.
We demonstrate the effectiveness of these guidelines by reviewing the use of qPCR to study anaerobic processes and biologic
nutrient removal processes. qPCR 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 ammonia-oxidizing
bacteria in various laboratory- and full-scale biologic processes. Researchers 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. 相似文献
3.
Kim Sangmin Kim Eunji Hwang Seokhwan 《Journal of Material Cycles and Waste Management》2022,24(6):2669-2676
Journal of Material Cycles and Waste Management - This study aims to investigate the methanogenic community of anaerobic mono-digestion of sewage sludge (SL-digester) and co-digestion of sewage... 相似文献
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