RNAseq of Pseudo-nitzschia arenysensis: Int11 day 12 rep 3
Wild type and transformants cultures were grown at 18C under white light at approximately 70 mol photons m-2 s-1 and 12:12h, dark: light cycle. Cells were collected at two time points of the growth curve in triplicate, at logarithmic phase T10 and at senescence phase T12. RNA was extracted following the manufacturers instructions (Trizol reagent, Invitrogen) and the genomic DNA contamination was removed by DNase I treatment (RNase-Free DNase Set Qiagen) followed by RNA purification using RNeasy Plant Mini Kit (Qiagen). The quantity of RNA was determined by Qubit assay (Qubit 2.0 Fluorometer, Life Technologies) and integrity was assessed by Bioanalyzer (2100 Bioanalyzer Instruments, Agilent Technologies) as well as by running samples on a 1.5% agarose gel. Libraries were prepared using 1.5 g RNA from each sample per time point with the Illumina TruSeq, following Manufacturers instructions. Libraries were sequenced on an Illumina HiSeq2000 (Single End 50 bp reads) at the GeneCore facility of the European Molecular Biology Laboratory (EMBL).