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Rapid and simple comparison of messenger rna levels using real-time PCR

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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Correspondence to Marc Pouliot.

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Open Access This article is published under license to BioMed Central Ltd. This is an Open Access article is distributed under the terms of the Creative Commons Attribution License ( https://creativecommons.org/licenses/by/2.0 ), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Dussault, AA., Pouliot, M. Rapid and simple comparison of messenger rna levels using real-time PCR. Biol. Proced. Online 8, 1–10 (2006). https://doi.org/10.1251/bpo114

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  • DOI: https://doi.org/10.1251/bpo114

Indexing terms

  • Polymerase Chain Reaction
  • Leukocytes
  • Gene Expression
  • RNA
  • Messenger