title: Illumina HiSeq 2500 paired end sequencing; Receptor Tyrosine Kinase pathway activation is sufficient to trigger chordoma in zebrafish
design: Receptor Tyrosine Kinase pathway activation is sufficient to trigger chordoma in zebrafish
library_construction_protocol: 8 dpf wildtype and Tg(twhh:Gal4);Tg(UAS:EGFP-HRASV12) were euthanized with 3% Tricaine methanesulfonate (Sigma). Embryos were decapitated and incubated in Tripsin-EDTA (Sigma) for 30 minutes to facilitate tissue dissociation. Notochords were then dissected using tungsten needles and immediately transferred to Trizol-LS (Ambion). We isolated 30-50 notochords per replicate, with a total of 3 replicates per condition (3x wildtype, 3x HRASV12). Notochord RNA was extracted following the manufacturer's protocol using Trizol-LS. RNA-seq libraries were constructed using the Truseq stranded total RNA kit.