title: RNA-seq of zebrafish embryos with mutations in a muscle-specific kinase and the giant titin protein
design: RNA-seq of zebrafish embryos with mutations in a muscle-specific kinase and the giant titin protein
library_construction_protocol: Zebrafish embryos were collected for DNA and RNA extraction at 5 dpf in 6 pools of 3 embryos per genotype. Samples were lysed in 110 μl RLT buffer (Qiagen) containing 1.1 μl of 14.3 M β-mercaptoethanol (Sigma). The lysate was allowed to bind to 450 μl of Agencourt AMPure XP beads (Beckman Coulter) for 15 minutes. The tubes were left on a magnet (Invitrogen) until the solutions cleared and the supernatant was then removed without disturbing the beads. Whilst still on the magnet, the beads were washed three times with 70% ethanol and allowed to dry for 20 minutes. Total nucleic acid was eluted from the beads following the manufacturer's instructions and treated with DNase I (NEB, Catalog number M0303L). NEBNext Ultra II DNA Library Prep Kit for Illumina.