Related Experiment Video
Updated: Aug 24, 2025
08:17
Cryosectioning of Contiguous Regions of a Single Mouse Skeletal Muscle for Gene Expression and Histological Analyses
Published on: December 12, 2016
15.9K
Comparison of reference gene expression stability in mouse skeletal muscle via five algorithms
Jianfeng Ma1,2,3, Jingyun Chen1,2,3, Mailin Gan1,2
1Department of Animal Science, College of Animal Science and Technology, Sichuan Agricultural University, Chengdu, China.
Peerj
|October 24, 2022
Summary
Selecting stable reference genes (RGs) is crucial for accurate gene expression analysis using real-time quantitative PCR (RT-qPCR). ACTB, HPRT, and YWHAZ are recommended for normalizing gene expression in mouse skeletal muscle studies.
Area of Science:
- Molecular Biology
- Genetics
- Biochemistry
Background:
- Real-time quantitative PCR (RT-qPCR) is vital for gene expression quantification.
- Reference gene (RG) selection is critical for reliable RT-qPCR results.
- Mouse skeletal muscle research requires validated normalization strategies.
Purpose of the Study:
- To identify the most stable reference genes for RT-qPCR in mouse skeletal muscle.
- To evaluate eight commonly used RGs across various experimental conditions.
- To provide a basis for accurate gene expression normalization in murine skeletal muscle.
Main Methods:
- RT-qPCR was used to assess the expression of eight candidate RGs (ACTB, GAPDH, HPRT, YWHAZ, B2M, PPIA, TUBA, 18S).
- Expression stability was analyzed using five computational programs: comparative ΔCq, NormFinder, BestKeeper, geNorm, and RefFinder.
- Analysis included diverse mouse skeletal muscle samples (genetic background, tissue type, growth stage) and C2C12 myoblasts.
Main Results:
- ACTB, HPRT, and YWHAZ demonstrated the highest expression stability.
- GAPDH and 18S were identified as the least stable reference genes.
- Normalization with ACTB, HPRT, and YWHAZ is recommended for mouse skeletal muscle RT-qPCR.
Conclusions:
- The study provides a validated set of reference genes for accurate gene expression studies in mouse skeletal muscle.
- Consistent normalization using ACTB, HPRT, and YWHAZ will enhance the reliability of RT-qPCR data.
- This research supports robust gene expression analysis in mammalian skeletal muscle research.
