Real‐time quantitative PCR: A tool for absolute and relative quantification
Real‐time quantitative PCR is a technique used to monitor the PCR reaction in real time. RT‐qPCR is broadly classified into two types based on its purpose: absolute and relative quantification. Absolute quantification is used in a wide array of fields such as microbiology, food technology, and biote...
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| Veröffentlicht in: | Biochemistry and molecular biology education 2021-09, Vol.49 (5), p.800-812 |
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| container_title | Biochemistry and molecular biology education |
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| creator | Harshitha, Ravikumar Arunraj, Duraipandian Rex |
| description | Real‐time quantitative PCR is a technique used to monitor the PCR reaction in real time. RT‐qPCR is broadly classified into two types based on its purpose: absolute and relative quantification. Absolute quantification is used in a wide array of fields such as microbiology, food technology, and biotechnology to quantify the microbiological load/adulterants in a commodity/copy numbers respectively, whereas Relative quantification is used in the field of genomics and functional transcriptomics to perform gene expression analysis in biological experiments. A laboratory work that covers the basic principles involved in RT‐qPCR and data analysis using the manual as well as the software methods are incorporated. The laboratory experiment was designed to provide insights on certain important principles such as primer characteristics, PCR efficiency, and melt curve analysis. This laboratory exercise provides all the significant components of RT‐qPCR, which can be useful while performing an experiment in an undergraduate and graduate laboratory. |
| doi_str_mv | 10.1002/bmb.21552 |
| format | Article |
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| ispartof | Biochemistry and molecular biology education, 2021-09, Vol.49 (5), p.800-812 |
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| language | eng |
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| source | Wiley Online Library Free Content; Access via Wiley Online Library; EZB-FREE-00999 freely available EZB journals; EBSCOhost Education Source; Alma/SFX Local Collection |
| subjects | absolute and relative quantification College Science copy number Data Analysis fold change Gene expression Laboratories Laboratory Experiments Laboratory Procedures Molecular Biology PCR efficiency quantitative real time PCR Science Instruction |
| title | Real‐time quantitative PCR: A tool for absolute and relative quantification |
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