GSM7995225: GFP positive H3K4me1 cells; Danio rerio; ChIP-Seq
1 µg of H3K4me1 antibody (ab8895, Abcam) was added to the cell lysates. Enhancer regions were incubated with magnetic Dynabeads™ Protein G (ThermoFisher Scientific) on a wheel at 4°C overnight. Beads have been washed for 10 min rotating on a wheel at 4°C with the following buffers: once with dilution buffer (165mM NaCl, 0.01% SDS, 1.1% Triton X-100, 1.2 mM EDTA pH 8.0, 16.7 mM Tris HCl pH 8.0), twice with low salt buffer (150 mM NaCl, 0.5% sodium deoxycholate, 0.1% SDS, 1% Nonidet P-40, 1 mM EDTA pH 8.0, 50 mM Tris HCl pH 8.0), twice with high salt buffer (500 mM NaCl, 0.5% sodium deoxycholate, 0.1% SDS, 1% Nonidet P-40, 1 mM EDTA pH 8.0, 50 mM Tris HCl pH 8.0), once with TE buffer (0.25 mM EDTA pH 8.0, 10 mM Tris HCl pH 8.0). Immunoprecipitated DNA was eluted by vortexing twice for 15 min at room temperature with 100 µl elution buffer (1% SDS, 100 mM sodium bicarbonate) each. DNA was de-crosslinked by adding NaCl to a final concentration of 0.3M with subsequent incubation at 65°C overnight, followed by purification (Qiagen MinElute PCR Purification Kit, #28004). Libraries have been prepared using the Clontech SMARTer ChIP-Seq Kit (Protocol 021115). A single equimolar pool was made based upon size-adjusted qPCR quantitation results, followed by denaturation of 14 pM used for c-bot hybridisation and cluster generation in an Illumina HiSeq 1500 High output run (Illumina Protocol 15006165 Rev K Oct 2012). Our samples were sequenced using 50 bp single reads (Illumina HiSeq High output Mode, Illumina Protocol 15035788 Rev D, Apr 2014).