Nampt hs00237184 m1

To quantify visfatin mRNA expression, a custom TaqMan™ gene expression assay (FAM/MGB probe; Life Technologies, USA) was performed using StepOnePlus™ real-time PCR system. NAMPT (Hs00237184_m1) and GAPDH as housekeeping (Hs99999905_m1) were purchased from Life technologies. A single qPCR reaction was performed in 20 µl volume containing 10 µl of 2x TaqMan gene expression master mix, 1 µl 20x TaqMan gene expression assay, 1 µl of cDNA sample, and 8 µl water free of RNase and DNase. Both targets and internal control were amplified in triplicate in all samples. The reactions were incubated at 50 °C for 2 min, activation step at 95 °C for 10 min, followed by 40 cycles at 92 °C for 15 s, 60 °C for 1 min. The visfatin expression was normalized to GAPDH, and relative expression was determined using the ΔΔCt method.

To quantify visfatin mRNA expression, a custom TaqMan™ gene expression assay (FAM/MGB probe; Life Technologies, USA) was performed using StepOnePlus™ real-time PCR system. NAMPT (Hs00237184_m1) and GAPDH as housekeeping (Hs99999905_m1) were purchased from Life technologies. A single qPCR reaction was performed in 20 µl volume containing 10 µl of 2x TaqMan gene expression master mix, 1 µl 20x TaqMan gene expression assay, 1 µl of cDNA sample, and 8 µl water free of RNase and DNase. Both targets and internal control were amplified in triplicate in all samples. The reactions were incubated at 50 °C for 2 min, activation step at 95 °C for 10 min, followed by 40 cycles at 92 °C for 15 s, 60 °C for 1 min. The visfatin expression was normalized to GAPDH, and relative expression was determined using the ΔΔCt method.