​Identification of Suitable Reference Genes for qPCR Analysis of 4T1 Mouse Mammary Tumor Cell Line

Abstract

Background: Identification of candidate reference genes for real time PCR study is a preliminary requirement to normalize experimental data and thus, deduce a reliable conclusion. Complex tissues like mouse mammary gland constitutes various cell types which makes it difficult to identify reference gene constantly expressing under different experimental conditions. Methods: In this study we have identified suitable reference genes for 4T1 tumor cell line derived from mouse mammary tumor cells. We have studied four genes namely Gapdh, Actb, Prdx1 and Ctbp1 for their expression stability in CPV2.NS1 post transfected 4T1 cells by Best Keeper. Result: By our study, three reference genes i.e. Prdx1, Gapdh and Ctbp1 were found to be quite correlated with the BestKeeper index, but by considering all three criteria of selection by BestKeeper algorithm, Prdx1 showed minimum standard deviation and coefficient of variation and was found to be ranked at first position by BestKeeper which suggests Prdx1 to be considered as better internal control gene among all other reference genes taken in our study for qPCR based experiments in 4T1 mouse mammary tumor cell line transfected with CPV2.NS1