Rapid and simple comparison of messenger rna levels using real-time PCR |
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Authors: | Andrée-Anne Dussault Marc Pouliot |
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Affiliation: | (1) Department of Anatomy-Physiology, Faculty of Medicine, Laval University, Quebec, Canada;(2) Centre de Recherche en Rhumatologie et Immunologie, CHUL, 2705 Laurier boulevard, Office T1-49, Sainte-Foy, G1V 4G2 Québec, Canada |
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Abstract: | Real-time polymerase chain reaction (PCR) constitutes a significant improvement over traditional end-point PCR, as it allows the quantification of starting amounts of nucleic acid templates, in real-time. However, quantification requires validation through numerous internal controls and standard curves. We describe in this paper a simple protocol which uses real-time PCR to compare mRNA levels of a gene of interest between different experimental conditions. Comparative real-time PCR can be a relatively low-cost method and does not require sequence-specific fluorescent reporters. Moreover, several genes from a set of experiments can be assessed in a single run. Thus, in addition to providing a comparative profile for the expression of a gene of interest, this method can also provide information regarding the relative abundance of different mRNA species. |
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Keywords: | KeywordHeading" >Indexing terms Polymerase Chain Reaction Leukocytes Gene Expression RNA Messenger |
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