GSM9169836: Fol4287 CTRL mono 3; Fusarium oxysporum; RNA-Seq
Germinating conidia / filaments were disrupted in a Minilys bead beater (Bertin Instruments, France) for 30 s at medium speed. Fractions from 3-12 and 13 -24 polysomes were combined and processed for RNA extraction using Trizol LS 3:1 ratio as the manufacturer's instructions.Total RNA was extracted with the Qiagen (USA) RNeasy micro kit and was treated on-column with RQ1 RNase-free DNase (Promega, USA) to remove additional residues of genomic DNA. RNA quality was quantified by Agilent 2200 TapeStation machine using an RNA kit (Agilent Technologies, USA). For quantification, 1 µl RNA was quantified with a Qubit 2.0 Fluorometer and RNA BR kit (Life Technologies, USA). cDNA libraries were prepared using 0.5 µg RNA with a QuantSeq 3ʹ mRNA-Seq Library Prep Kit FWD (Lexogen GmbH, Austria) and were sequenced using the NextSeq 500 system (Illumina, USA) at the Center for Genomic Technologies of the Hebrew University of Jerusalem.