GSM1348923: Af_30min-O2-B3; Aspergillus fumigatus; RNA-Seq
Culture aliquots of 200 ml were taken before and at indicated time points throughout the shift. The mycelium was rapidly separated from the medium by filtration, immediately shock frozen in liquid N2 and stored at -80°C until RNA isolation. A phenol-chloroform based method was used to extract total RNA from mycelia. First, 100-mg-aliquots of the deeply frozen mycelium of A. fumigatus were mechanically disintegrated in the presence of 1 ml of TriSureTM (Bioline GmbH, Luckenwalde, Germany) using the FastPrep®24 instrument (MP Biomedicals, Illkirch, France). Following the addition of chloroform and centrifugation, RNA was precipitated from the aqueous phase and solubilized in ultrapure H2O.RNA was e additionally digested with DNaseI and column purified using the RNeasy spin columns (Qiagen, Hilden, Germany) to remove DNA and phenol contaminations, respectively. Concentration and integrity of the RNA were checked by Nanodrop measurements and agarose gel electrophoresis. tagged random primed cDNA libraries were prepared from fragmented polyA-RNA using standard illumina protocols
GEO Accession
GSM1348923