Illumina HiSeq 2500 paired end sequencing; Neochloris oleoabundans under salt and nitrogen stress
Neochloris oleoabundans under salt and nitrogen stress
Fresh water with nitrogen Samples for RNA isolation were immediately processed after sampling and kept on ice. Cells were collected by centrifugation at 4.000xg at 0°C for three minutes and immediately frozen in liquid nitrogen before storage at -80°C until further extraction. Cells were disrupted by grinding the pellet using mortar and pestle and liquid nitrogen. A 5 ml volume of heated (65°C) phenol-chloroform and 5 ml of extraction buffer (10 mM EDTA, 1% sodium dodecyl sulphate (SDS), 2% 2-mercaptoethaol and 200 mM sodium acetate, pH 5) was added to the ground biomass. The RNA was precipitated by addition of 1/3 volume 8M lithium chloride (LiCl) enriched with 1% 2-mercaptoethanol. The RNA pellet was washed with 2M LiCl and twice with 70% ethanol. After evaporation of the last residues of ethanol, the pellet was resuspended in RNase free H2O. Illumina TruSeq RNA protocol
Experimental Factor: Fresh water, nitrogen replete
growth condition