GSM5123792: 2 dpt inf R3; Arabidopsis thaliana; RNA-Seq
For Fo5176 transcriptomics in the absence of a plant host, 107 microconidia/ml were germinated in ½ MS + 1% (w/v) sucrose overnight at 180rpm at 28°C in the dark. The germinated microconidia were harvested via two centrifugation steps at 4000 xg for 15 minutes at 10° C, washed twice with water, discarding supernatant in between washes. 3'mRNA-libraries were prepared using the 3'mRNA-Seq library Prep Kit (QuantSeq, Lexogen). The manual from the supplier was followed with the following modifications. At least 1 µg RNA was used as input. For first strand synthesis of cDNA the incubation time at step 4 was increased to 60 min. For library indexing and amplification 13-17 amplification cycles of the given PCR were used depending on the amount of input RNA. 17 µl of the purified library was transferred to a fresh tube. The finished libraries were stored at -20° C until quality control and pooling.