Objective:To develop the method of detecting the level ofHey1 expression_the target gene of Notch signaling pathway in bone marrow mesenchymal stem cells

using real-time fluorescence quantitative RT-PCR.Methods:The bone marrow mesenchymal stem cells (C2C12) were cultured and the total RNA was extracted.And then the amplification efficiency was verified between the reference gene β-actin and the target gene Hey1.To optimize the PCR conditions and analyse amplification curves and melting curve before validation of quantitatve PCR products with gel electrophoresis.Results:The reference geneβ-actin and the target gene Hey1 have the same amplification efficiency(slope difference of the amplification curve<0.1)

suggesting that you can use the comparative Ct method for analysing relative expression of Hey1.Through analysing the amplification curve

melting curve and gel electrophoresis results we can get that the qPCR product was very specific.Conclusion:The real-time quantitative RT-PCR methods established for detecting the expression of Hey1 gene is accurate and reliable

which can be used for studying the relationship of Bmp9 and Notch signaling pathways.