design: RNA-Seq of zebrafish embryos (96hpf) treated with different concentrations of Methimazole against untreated control groups
library_construction_protocol: at the end of exposure, 10 Embryos were euthanized on ice then homogenised in lysis buffer using Lysing Matrix D ceramic beads in FastPrep-24 homogeniser (MP Biomedicals, Irvine, USA) at 5 m/s speed for 1 min. Zebrafish embryos were exposed to Methimazole (CAS 60-56-0), for 96 hours according to OECD test guidelines No. 236 at 27 ± 1 °C and a light/dark cycle of 14:10 hours. three zebrafish embryos replicate groups originating from three different tanks were exposed to each of two sub-lethal concentrations of methimazole (80 and 160 mg/L) in addition to untreated control groups. At the end of exposure time (96 hours), RNA was extracted from 10 embryos using a NucleoSpin RNA/Protein kit (Macherey-Nagel, Düren, Germany). RNA libraries were prepared by the Eurofins Genomics Germany GmbH from RNA extracts with RNA integrity number (RIN) values > 9 using poly(A) RNA purification followed by fragmentation. random-primed strand-specific cDNA libraries were then prepared.