MilliporeSigma
  • Home
  • Search Results
  • Evaluation of reference genes for quantitative PCR analysis of mouse lymphocytes.

Evaluation of reference genes for quantitative PCR analysis of mouse lymphocytes.

Journal of immunological methods (2012-08-14)
Tina C Albershardt, Brian M Iritani, Alanna Ruddell
ABSTRACT

Normalization to a reference gene is the method of choice for quantitative PCR analysis. The stability of reference genes is critical for accurate gene expression analysis, as significant variations in reference gene expression can alter experimental results and conclusions. In this study, we evaluated the expression stability of five commonly used reference genes found in mouse lymphocytes. Using NormFinder and BestKeeper algorithms, we consistently show that ubiquitin C (Ubc) is the optimal reference gene for normalizing qPCR data obtained from mouse lymphocytes, whereas beta-actin (Actb) is not a suitable reference gene due to its extensive variability in expression. Our findings emphasize the importance of validating reference genes for qPCR analyses. We provide a shortlist of reference genes to use for normalization and recommend freely available software programs as a rapid approach to validate potential reference genes.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Glyceraldehyde-3-phosphate Dehydrogenase from rabbit muscle, lyophilized powder, ≥75 units/mg protein
Supelco
GAPDH, standard for protein electrophoresis
Sigma-Aldrich
Glyceraldehyde-3-phosphate Dehydrogenase from human erythrocytes, lyophilized powder, 50-150 units/mg protein