4.6. RNA Extraction and cDNA Synthesis

For the analysis of gene expression, the fungal mycelium was first washed in a cold Phosphate-Buffered Saline (PBS) buffer and then dried in a freezer-dryer. The freeze-dried mycelium was used. Extraction of RNA from the fungal mycelium was performed using TRIzol^® reagent (Invitrogen, Carlsbad, CA, USA) and according to the manufacturer’s instructions. Samples were treated with DNase I. To quantify RNA, a NanoDrop OneC Spectrophotometer (Thermo Fisher Scientific, Waltham, MA, USA) at a wavelength of 260 and 280 nm was used. To determine the quality of the extracted RNA, electrophoresis was performed using agarose gel (1.2% agarose). Total purified RNA was stored at −80 °C until further processing. Via reverse transcription, the cDNA was synthesized from 0.6–2 μg of RNA and random primers by using a RT-PCR kit (SensiFAST™ cDNA Synthesis Kit, Bioline Reagents, Ltd., London, UK).