Molecular Neurobiology

, Volume 53, Issue 3, pp 1540–1550 | Cite as

Normalization of Reverse Transcription Quantitative PCR Data During Ageing in Distinct Cerebral Structures

  • G. Bruckert
  • D. Vivien
  • F. Docagne
  • B. D. Roussel
Article

Abstract

Reverse transcription quantitative-polymerase chain reaction (RT-qPCR) has become a routine method in many laboratories. Normalization of data from experimental conditions is critical for data processing and is usually achieved by the use of a single reference gene. Nevertheless, as pointed by the Minimum Information for Publication of Quantitative Real-Time PCR Experiments (MIQE) guidelines, several reference genes should be used for reliable normalization. Ageing is a physiological process that results in a decline of many expressed genes. Reliable normalization of RT-qPCR data becomes crucial when studying ageing. Here, we propose a RT-qPCR study from four mouse brain regions (cortex, hippocampus, striatum and cerebellum) at different ages (from 8 weeks to 22 months) in which we studied the expression of nine commonly used reference genes. With the use of two different algorithms, we found that all brain structures need at least two genes for a good normalization step. We propose specific pairs of gene for efficient data normalization in the four brain regions studied. These results underline the importance of reliable reference genes for specific brain regions in ageing.

Keywords

RT-qPCR Ageing Normalization geNorm Normfinder 

Notes

Acknowledgments

We would like to thank the Basse-Normandie region for funding Bruckert G and Dr Roussel BD and its credits for the laboratory.

Supplementary material

12035_2015_9114_MOESM1_ESM.docx (24 kb)
Supplementary table 1An intragroup analysis was performed using the Mann and Whitney non-parametric test (Grap Prism software) on the raw data for each genes studied in the cortex, hippocampus, striatum and cerebellum. *: p < 0.05; **: p < 0.01; ***: p < 0.001. (DOCX 23 kb)

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • G. Bruckert
    • 1
  • D. Vivien
    • 1
  • F. Docagne
    • 1
  • B. D. Roussel
    • 1
  1. 1.INSERM, INSERM U919, Serine Proteases and Pathophysiology of the Neurovascular Unit, GIP CYCERONUniversity Caen Basse NormandieCaenFrance

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