Journal article
Validation of reference genes for normalization of real-time quantitative RT-PCR data in traumatic brain injury
Journal of Neuroscience Research, Vol.87(1), pp.34-41
2009
Abstract
Careful validation of reference genes used for the normalization of real-time RT-PCR data is required to obtain accurate results regarding gene expression. We evaluated the stability of seven commonly used reference genes in the cerebral cortex and hippocampus of rats 3 days following traumatic brain injury (TBI). HPRT, SDHA, and GUSB were found to be the most stable reference genes in the cerebral cortex, whereas B2MG, TBP, and GAPDH were the most stable in the hippocampus. The use of three reference genes was determined to be the optimal number for accurate normalization of data. To illustrate this point, when our gene of interest, substance P (SP), was normalized against the three most stable reference genes in both brain areas, we found no significant difference between injured and uninjured rats at the 3-day time point. However, when our SP data were normalized to each reference gene individually, SP mRNA level was highly variable depending on the reference gene chosen. The results of the present study highlight the importance of validating reference genes to be used for real-time RT-PCR analysis. The use of the most stable reference genes presented here will allow more accurate normalization of gene expression data in TBI. © 2008 Wiley-Liss, Inc.
Details
- Title
- Validation of reference genes for normalization of real-time quantitative RT-PCR data in traumatic brain injury
- Authors
- N L Cook (Author) - University of AdelaideR Vink (Author) - University of AdelaideJames J Donkin (Author) - University of AdelaideC van den Heuvel (Author) - University of Adelaide
- Publication details
- Journal of Neuroscience Research, Vol.87(1), pp.34-41
- Publisher
- John Wiley & Sons Inc.
- Date published
- 2009
- DOI
- 10.1002/jnr.21846
- ISSN
- 0360-4012
- Organisation Unit
- UniSC Clinical Trials Centre; University of the Sunshine Coast, Queensland
- Language
- English
- Record Identifier
- 99450426102621
- Output Type
- Journal article
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