mRNA-Seq of Danio rerio exposed to different concentrations of Fulvestrant against untreated control groups
mRNA-Seq of Danio rerio exposed to different concentrations of Fulvestrant against untreated control groups
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 45 seconds. Zebrafish embryos were exposed to Fulvestrant 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 (EC10 and EC5) of Fulvestrant (CAS 129453-61-8) in addition to untreated control groups. total RNA and Protein were extracted from the tissue lysate using using the NucleoSpin® RNA/Protein kit (Macherey & Nagel, Düren, Germany). according to the manufacturer's protocol. Nanodrop 2000 spectrophotometer (Thermo Scientific) was used to measure and normalize the obtained RNA concentrations. The quality of the extracted RNA samples was checked using RNA 6000 Nano kit in 2100 Bioanalyzer System (Agilent, Santa Clara, USA). Samples were stored at -80°C.    RNA libraries were prepared by the Eurofins Genomics Germany GmbH from RNA extracts with RNA integrity number (RIN) values > 7 using poly(A) RNA purification followed by fragmentation. Using reverse transcription, cDNA libraries was produced from mRNA fragments and ligated to the adapters.