RNA-seq of E.huxleyi life phases
RNA was extracted from cell pellets using the RNeasy Plant Kit (Qiagen) and treated with DNase (Turbo DNA-free kit, Ambion). RNA concentration and integrity were measured by Nanodrop (Thermo Fisher Scientific) and Tapestation (Agilent Genomics). cDNA libraries were prepared according to the Illumina RNA sample preparation kit. mRNA was isolated using poly(T) oligo-attached magnetic beads, fragmented into 200-500 bp pieces and RNA fragments were reverse transcribed with SuperScript II reverse transcriptase (Life Technology). Adapters ligation, amplification and purifications preceded the sequencing of the cDNA libraries on an Illumina NextSeq platform into 75-bp single-end reads.