RNA-seq of zebrafish: BPA_2
Total RNA was isolated from single embryos at 24 hpf using NucleoSpin RNA XS (Macherey-Nagel). After quality assessment of RNA using NanoDrop (Thermo Fisher Scientific), Picopgreen (Invitrogen), and Agilent 2100 Bioanalyzer, high-quality samples were used to construct libraries using the SMARTer Ultra low input RNA kit. Libraries were sequenced with paired ends using the Illumina HiSeq 2500 platform at Macrogen (Tokyo, Japan). RNA isolation, cDNA library construction, and sequencing were performed twice, using three embryos per experimental condition. Samples 13 and 79 were treated separately for RNA isolation and sequencing but analyzed simultaneously.