GSM9155750: Section D 1 hpf rep 3; Danio rerio; RNA-Seq
The samples were homogenized in 300 μl of TRIReagent® (Sigma-Aldrich, USA) and total RNA was extracted according to the manufacturer's protocol. LiCl precipitation was performed to remove inhibitors present in the yolky vegetal hemisphere. The concentration of RNA was measured using NanoDrop-2000 (ThermoFisher, USA) and sample quality was assessed using 5200 Fragment Analyzer (Agilent, USA). Libraries were prepared using NEBNext® Ultra™ II Directional RNA Library Prep Kit for Illumina® (New England Biolabs, USA) and adaptors and indexes were provided with NEBNext® Multiplex Oligos for Illumina® (96 Index Primers) (New England Biolabs, USA). 20 ng of a total RNA was used for library preparation. Ribosomal RNA depletion was performed using Ribocop rRNA Depletion Kit V1.3 (Lexogen, Austria).