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  • Open Access

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

Biological Procedures Online8:81001

  • Received: 23 November 2005
  • Accepted: 16 December 2005


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.

Indexing terms

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