GSM8119571: QM9414 SESp-cbh1 mutXYR1, batch lactose day 2, rep2; Trichoderma reesei; RNA-Seq
Total RNA was extracted from mycelia using TRIzol (Thermo Fisher Scientific) according to manufacturer's instructions with small modifications. Mycelium samples were ground using cooled mortars under liquid nitrogen into fine powder and about 50 µL of powder was transferred into a 2 mL tube, after which 1 mL of TRIzol was immediately added. Mixture was vortexed and incubated at room temperature for 5 minutes, after which it was centrifuged (12000 x g, 4 °C, 10 minutes) and the supernatant was transferred to a new tube. After this, the protocol was continued according to manufacturer's instructions. RNA was then purified using RNeasy Mini kit (Qiagen) according to manufacturer's instructions, except that 950 µl of 94 vol-% ethanol was added instead of 250 µl during the second step. Extracted and purified RNA samples were sent for RNA-sequencing in Source BioScience (Cambridge, UK). rRNA depletion libraries were prepared using Next Ultra II Directional RNA Library Prep kit (New England Biolabs). Sequencing of the rRNA libraries was done using Illumina NovaSeq 6000.