Reverse transcription
Extracted RNA was reverse transcribed using high-capacity cDNA reverse transcription kit, Thermo Fisher Scientific, USA. Extracted RNA (1µg diluted to the correct volume 10 µl by Nuclease- free H2O) were added to 10 μL of 2X RT master mix. The 2X RT master mix contained 2 µL 10X RT Buffer, 0.8 µL 25X dNTP Mix (100 mM), 2 µL 10X RT Random Primers, 1 µL MultiScribe™ Reverse Transcriptase, 1 µL RNase Inhibitor 3.2 µL Nuclease-free H2O. The thermal cycler; Biorad, USA, program was 25°C for 10 minutes, 37°C for 120 minutes, 85°C for 5 minutes and 4°C for∞.