PUBLICATION
Expressed repetitive elements are broadly applicable reference targets for normalization of reverse transcription-qPCR data in mice
- Authors
- Renard, M., Vanhauwaert, S., Vanhomwegen, M., Rihani, A., Vandamme, N., Goossens, S., Berx, G., Van Vlierberghe, P., Haigh, J.J., Decaesteker, B., Van Laere, J., Lambertz, I., Speleman, F., Vandesompele, J., Willaert, A.
- ID
- ZDB-PUB-180518-4
- Date
- 2018
- Source
- Scientific Reports 8: 7642 (Journal)
- Registered Authors
- Speleman, Frank, Willaert, Andy
- Keywords
- none
- MeSH Terms
-
- Animals
- Cardiovascular Abnormalities/genetics*
- Cardiovascular Abnormalities/pathology
- Disease Models, Animal
- Female
- Humans
- Leukemia, Prolymphocytic, T-Cell/genetics*
- Leukemia, Prolymphocytic, T-Cell/pathology
- Male
- Mice
- Mice, Inbred C57BL
- Mice, Transgenic
- Neuroblastoma/genetics*
- Neuroblastoma/pathology
- Reference Standards
- Repetitive Sequences, Nucleic Acid/genetics*
- Reverse Transcriptase Polymerase Chain Reaction/methods
- Reverse Transcriptase Polymerase Chain Reaction/standards*
- Skin Diseases/genetics*
- Skin Diseases/pathology
- Zebrafish/metabolism
- PubMed
- 29769563 Full text @ Sci. Rep.
Citation
Renard, M., Vanhauwaert, S., Vanhomwegen, M., Rihani, A., Vandamme, N., Goossens, S., Berx, G., Van Vlierberghe, P., Haigh, J.J., Decaesteker, B., Van Laere, J., Lambertz, I., Speleman, F., Vandesompele, J., Willaert, A. (2018) Expressed repetitive elements are broadly applicable reference targets for normalization of reverse transcription-qPCR data in mice. Scientific Reports. 8:7642.
Abstract
Reverse transcription quantitative PCR (RT-qPCR) is the gold standard method for gene expression analysis on mRNA level. To remove experimental variation, expression levels of the gene of interest are typically normalized to the expression level of stably expressed endogenous reference genes. Identifying suitable reference genes and determining the optimal number of reference genes should precede each quantification study. Popular reference genes are not necessarily stably expressed in the examined conditions, possibly leading to inaccurate results. Stably and universally expressed repetitive elements (ERE) have previously been shown to be an excellent alternative for normalization using classic reference genes in human and zebrafish samples. Here, we confirm that in mouse tissues, EREs are broadly applicable reference targets for RT-qPCR normalization, provided that the RNA samples undergo a thorough DNase treatment. We identified Orr1a0, Rltr2aiap, and Rltr13a3 as the most stably expressed mouse EREs across six different experimental conditions. Therefore, we propose this set of ERE reference targets as good candidates for normalization of RT-qPCR data in a plethora of conditions. The identification of widely applicable stable mouse RT-qPCR reference targets for normalization has great potential to facilitate future murine gene expression studies and improve the validity of RT-qPCR data.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping