Lightcycler 480 dna sybr green 1 master chemistry

RNA was isolated from 2 × 10⁵ cells using the RNeasy 96 Mini Kit (Qiagen, Hiden, Germany) and converted to cDNA by the SuperScript III First-Strand Synthesis SuperMix kit (Invitrogen, Carlsbad, CA). Levels of STAT3 and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) transcripts were determined by real-time polymerase chain reaction (RT-PCR) using LightCycler 480 DNA SYBR Green I Master chemistry on LightCycler 480 instrument (Roche, Basel, Switzerland). Results were expressed as the fold change versus the result for the corresponding GAPDH housekeeping gene mRNA values using the 2-ΔΔCt method.³¹ (link)

RNA was isolated from 2 × 10⁵ cells using RNeasy 96 Mini Kit (Qiagen, Hilden, Germany), converted to complementary DNA by SuperScript III First-Strand Synthesis SuperMix kit (Invitrogen, Carlsbad, CA). Levels of STAT1 and GAPDH transcripts were determined by real-time quantitative polymerase chain reaction using LightCycler 480 DNA SYBR Green I Master chemistry on LightCycler 480 instrument (Roche, Basel, Switzerland), as reported before.²⁰ (link) The primer sets for STAT1 and GAPDH genes are provided in the Supplementary Table S1. Results were expressed as the fold change versus the control groups after the results were normalized with corresponding GAPDH housekeeping gene mRNA values using the 2^–ΔΔCT method.²⁸ (link)