title: ChIP-seq of zebrafish whole embryos
design: For Gfi1aa-eGFP ChIP, ~250 embryos injected with pTol2-hsp70:gfi1aa-eGFP construct were raised to 12 hpf for 2 h heat-shock treatment at 39 and recovered for 1 hour, and then fixed by 1.1% formaldehyde at 15 hpf. Then cross-linked embryos were lysed and sonicated by Covaris M220 sonicator (duty factor 20%, peak incident power 75, cycles of burst 200, time = 10 min) to a size range from 200 to 700 base pairs. Then sonicated chromatin was incubated together with Dynal Protein G magnetic beads and 5 ug antibody. Finally, after washed and cross-linking reversed, DNA was purified by phenol-chloroform-isoamyl alcohol. Then libraries were prepared using the NEBNext Ultra II DNA Library Prep Kit for Illumina and sequenced on the Illumina NovaSeq6000 sequencer with PE150 strategy.