4.6. RNA Isolation, cDNA Synthesis and Quantitative Real-Time PCR

Total mRNA was isolated using the RNeasy Mini Kit (Qiagen, Hilden, Germany) according to the manufacturer’s protocol. cDNA synthesis was performed using the QuantiTect Reverse Transcription Kit (Qiagen, Hilden, Germany) with an extended incubation time of 30 min at 42 °C. qRT-PCR was performed using the QuantiTect SYBR Green RT-PCR Kit (Qiagen, Hilden, Germany) and self-designed primers (Table S2) on the Lightcycler 96 instrument (Roche, Mannheim, Germany). All values were adjusted to TBP (TATA-box binding protein) as a reference gene.