GSM7867713: Dicty, whole organism, 2h, rep2; Dictyostelium discoideum; RNA-Seq
108 of cells from fresh culture (1-2 days, 0.5-1×106 cells per ml) or from development plates were harvested for 5 minutes at 400 g (room temperature), washed with 5 ml of 1× KK2 buffer and snap-frozen in a liquid nitrogen. RNA extraction was carried out using an RNeasy Mini kit (Qiagen) following the manufacturer's instructions. RNA quality was assessed using capillary electrophoresis with a Bioanalyzer 2100 (Agilent). For library preparation, a TruSeq RNA Sample Prep kit v2 (Illumina) was used following the manufacturer's instructions. After preparation, libraries were quantified using a Qubit fluorometer and quantitative PCR and sequenced with HiSeq 2000 with read lengths of 51 nt.