GSM4627066: SNPC13ChIP_N2_48h_IP_rep2; Caenorhabditis elegans; ChIP-Seq
After washing, the gut was cleared for 15 min by nutation in M9, followed by three washes in M9. Worms were live-crosslinked in 2.6% formaldehyde in water for 30 min at room temperature, nutating. Crosslinking was quenched with a final concentration of 125 mM glycine for 5 min with nutation. After three washes with water, worms were flash-frozen in liquid nitrogen. Frozen worms were ground into powder using a mixer mill MM400 (Retsch) for 2 rounds of 1 min at 30-1. Powder was resuspended in 1x RIPA buffer (1xPBS, 1% NP-40, 0.5% sodium deoxycholate, 0.1% SDS) for 10 min at 4°C. Crosslinked chromatin was sonicated using a Diagenode Bioruptor Pico for three 3-min cycles, 30 sec on/off. 10 µg chromatin was nutated overnight at 4°C with 2 µg of Flag antibody (Sigma-Aldrich, F1804) and then for 1.5 h with 50 µL mouse IgG Dynabeads (Invitrogen). Input amount was 10% of IP. Chromatin was decrosslinked and extracted as described previously (Weiser et al., 2017). Libraries were prepared and multiplexed using the Ovation Ultralow Library Systems v2 (NuGEN Technologies) according to the manufacturer's protocol.  The Illumina HiSeq 4000 platform was used to generate 50 bp single-end reads.
GEO Accession
GSM4627066