GSM5255621: wfikkn1-mut-DMSO-rep1_S3; Danio rerio; RNA-Seq
Total RNA was extracted from 48 hpf whole embryos using RNAzol (Molecular Research Center, Inc; Cincinnati, OH) and a bullet blender with 0.5 mM zirconium oxide beads (Next Advance, Averill Park, New York) as recommended by the manufacturer. The RNA was purified using the Direct-zol MiniPrep kit (Zymo Research; Irvine, CA) and included an in-column DNase 1 digestion. RNA quality and quantity was assessed using a SynergyMix microplate reader with the Gen5 Take3 module. RNA integrity was confirmed (RIN score > 8) with an Agilent Bioanalyzer 2100. Total RNA samples were sent to Oregon State University Center for Genome Research and Biocomputing Core facilities for library preparation and sequencing. Samples were prepared using the Robotic PolyA Enrichment Library Prep and Robotic Stranded RNA Library Prep Kits (WaferGen, Fremont, CA). Libraries were randomized across 6 lanes and sequenced with 100bp single-end reads using the Illumina HiSeq 3000. Raw sequence reads were quality checked prior to processing.