title: GSM3045712: Sibling rep2; Danio rerio; RNA-Seq
library_construction_protocol: For isolating endothelial cells from Tg(kdrl:EGFP)s843 siblings and Tg(kdrl:EGFP)s843 hhex mutant at 48 hpf, embryos were divided based on the absence of PLs.  Then, embryos were rinsed in HBSS (Gibco) followed by dissociation in TrypLE express (Gibco) at 28°C with repeated pipetting. Endothelial cells were isolated using BD Aria sorter and sorted for EGFP+ or tdTomato+/EGFP+ signals (for Tg(kdrl:EGFP)s843 Tg(fli1a:tdTomato-2A-hhex)bns136). RNA was isolated using the miRNeasy micro Kit (Qiagen) combined with on-column DNase digestion (DNase-Free DNase Set, Qiagen) to avoid contamination by genomic DNA. 6ng total RNA was used as input for SMARTer® Stranded Total RNA-Seq Kit - Pico Input Mammalian Kit (Clontech). Sequencing was performed on the NextSeq500 instrument (Illumina) using v2 chemistry, resulting in minimum of 30M reads per library with 1x75bp single end setup.