RNA-seq of Trebouxia lynnae: several physiological treatments
An equimolar mixture of the RNA extracted from the nine conditions was used to prepare the library for sequencing. Any rRNA was removed using Epicentre ribo-depletion protocol with a Ribo-Zero rRNA Removal Kit Plant Leaf (Illumina Inc., San Diego, California, USA). The ribo-depleted RNA was fragmented by incubation for 5 min at 94C, the library was constructed using the Illumina TruSeq stranded mRNA library preparation kit following the manufacturer's instructions. It was subjected to normalization following Illumina's DSN Normalization protocol prior sequencing. Sequencing was carried out in a Illumina MiSeq benchtop sequencer with a v3 2x200 pb kit at the Sequencing Service of the Servicio Central de Soporte a la Investigacin Experimental (SCSIE) of the University of Valencia (Burjassot, Spain).