GSM9067004: GS-C-VA3_2; Gelatoporia subvermispora; RNA-Seq
Transcriptomic samples were generated by centrifugation at 6,000 x g for 10 min at 4°C, followed by flash freezing the pellets in liquid nitrogen after discarding the supernatants. Samples were stored at -80°C. Frozen fungal cultures (100-200 mg) were transferred to a pre-chilled mortar and ground to a fine powder in liquid nitrogen. The liquid nitrogen was allowed to evaporate and the sample was transferred to a pre-chilled 15 mL centrifuge tube. All steps were performed on dry ice to ensure samples remained frozen. RLT buffer (700 μL)(Qiagen) containing 10% β- mercaptoethanol was added to the frozen samples and the samples were allowed to thaw with periodic vortexing at high speed. Samples were vortexed at high speed continuously for 1 min or until the sample was completely homogenous. Samples were transferred to a QIAshredder column (Qiagen) and processed using the RNeasy mini kit and subjected to DNase gDNA digestion according to the manufacturers protocol (Qiagen). The RNA samples are assessed using the Agilent 2100 Bioanalyzer, RNA with RIN >7 proceed with cDNA library. TruSeq Stranded Total RNA (cat#20020596) is used to generate cDNA library for illumina NextSeq550 platform according to the manufacture protocol. Single- read sequencing of the cDNA libraries with a read length of 150 is performed on NextSeq 500 Sequencing System using NextSeq 500/550 High Output v2 kit 150 cycles (cat#20024907).
GEO
GSM9067004_r1