GSM5211540: 23h input; Caenorhabditis elegans; ChIP-Seq
worms were incubated in M9 with 2% Formaldehyde for 30 minutes at room temperature with gentle agitation to allow protein-DNA crosslinking. Worms were lysed with beads using the FastPrep-24 5G machine (MP Biomedicals, settings: 8 m/sec, 30 sec on, 90 sec off, 5 cycles). Lysates were sonicated using the Bioruptor Plus Sonication system (Diagenode, settings: 30 sec on, 30 sec off, 20 cycles). 250 μg sonicated chromatin was incubated with 10 μg mouse anti-RNA polymerase II CTD antibody (8WG16, Abcam) at 4°C for 2 hours with gentle agitation and subsequently with 45 μL Dynabeads Protein G (Thermo Fisher Scientific) at 4°C overnight with gentle agitation. Elute was treated with 0.13 ug/uL RNase and 1 ug/uL Proteinase K. ChIP-seq libraries were prepared using NEBNext Ultra DNA Library Prep Kit for Illumina (New England Biolabs)