Real-time quantitative reverse transcription polymerase chain reaction (real-time qRT-PCR) was carried out with the Single Tube Custom TaqMan Gene Expression Assays (Thermo Fisher Scientific) and the LightCycler1 480 Probes Master (Roche Applied Science) using the Light Cycler 480 II detection system (Roche Applied Science). Expression values were normalized against control genes: GAPDH and TBP. The TaqMan probes used were as follows: ASAH (Hs00602774_m1), GAPDH (Hs02786624_g1), GLA (Hs00609238_m1), GM2A (Hs00166197_m1), HPSE (Hs00935036_m1), HYAL4 (Hs00202177_m1), IVL (Hs00846307_s1), LAMP1 (Hs00931461_m1), MCOLN1 (Hs01100653_m1), MITF (Hs01117294_m1), MTORC1 (Hs00234508_m1), PPP3CA (Hs00174223_m1), PPP3CB (Hs00236113_m1), PSAP (Hs01551096_m1), S100A7 (Hs01923188_u1), S100A9 (Hs00610058_m1), SMPD1 (Hs03679346_g1), SPHK1 (Hs00184211_m1), TBP (Hs00427620_m1), TFE3 (Hs00232406_m1), TFEB (Hs00292981_m1), and TFEC (Hs00992838_m1). Each experiment of real-time qRT-PCR analysis was repeated at least three times (n). In case of HaCaT cells they were biological replications and data are reported as the mean SD with p < 0.05 considered statistically significant. For skin specimens technical repetitions were conducted.
Single tube custom taqman gene expression assays
Manufactured by Thermo Fisher Scientific
Sourced in United States
Single Tube Custom TaqMan Gene Expression Assays are pre-designed assays for the quantitative analysis of gene expression. These assays use TaqMan technology to provide specific and sensitive detection of target gene sequences.
Automatically generated - may contain errors
2 protocols using single tube custom taqman gene expression assays
1
Real-time qRT-PCR Analysis of Skin Biomarkers
2
Real-Time qRT-PCR Analysis of Immune-Related Genes
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Analysis was carried out with Single Tube Custom TaqMan Gene Expression Assays (Thermo Fisher Scientific, Waltham, MA, USA) and the LightCycler 480 Probes Master (Roche Applied Science, Penzberg, Germany) using the Light Cycler 480 II detection system (Roche Applied Science, Penzberg, Germany). Expression values were normalised against control gene TATA-Box Binding Protein (TBP). The TaqMan probes used were as follows: CAMP (Hs00189038_m1), CCL20 (Hs00355476_m1), DEFB4A/B (Hs00823638_m1), MTORC1 (Hs00234508_m1), PIK3CA (Hs00907957_m1), S100A7 (Hs01923188_u1), S100A9 (Hs00610058_m1), TBP (Hs00427620_m1), and TFEB (Hs00292981_m1). Each experiment of real-time qRT-PCR analysis was repeated triple (n = 3). Data are reported as the mean ± standard deviation (SD) with p < 0.05 considered statistically significant.
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