title: GSM5097168: 4hpf_input; Danio rerio; ChIP-Seq
library_construction_protocol: Briefly, approximately 200-300 embryos were dechorionated with pronase and deyolked with deyolking buffer (55mM NaCL, 1.8mM KCl, 1.25mM Sodium Biocarbonate). Zebrafish cells were fixed using formaldehyde and stopped with 0.2M  glycine. Cells were snap frozen with liquid nitrogen and stored at -80C. Approximately 1000 embryos were used for the 4hpf Chip-seq was fragmented to a size range of 200–700bp with a Branson 250 Sonifier. Solubilized chromatin was immunoprecipitated with antibody.  Protein A‐sepharose was used to pull‐down the antibody–chromatin complexes which were then washed and eluted. After cross‐link reversal (5 hours, 65C) and proteinase K digestion, immunoprecipitated DNA was extracted with phenol‐chloroform, ethanol precipitated, and treated with RNase. NEBNext ChIP-Seq Library Prep Master Mix Set for Illumina (E6240) with NEBNext Multiplex Oligos for Illumina (E7335L)