GSM8777160: ChIPseq_NHR23_IP; Caenorhabditis elegans; ChIP-Seq
5 million animals were collected in PBS + PI + PMSF 1mM at 20 hours after plating and popcorn was created from resuspended larvae mix. Lysate was prepared by grinding the popcorn, crosslinking with 1.1% formaldehyde and sonication with a Diagnode Biorupter Pico. For the chromatin immuno-precipitation, 50 uL of protein G Dynabeads (10003D, ThermoFisher Scientific) were incubated with 5 ug of anti-GFP antibody (ab290, Abcam) for 4 hours rotating at 4°C. 100 ug of chromatin was added, and incubated over night at 4°C rotating. Beads were washed and the elute was treated with proteinase K and RNaseA, and DNA was purified using Zymo ChIP Concentrator Kit. Sequencing libraries were prepared using the ChIP-seq NEB Ultra protocol (New England Biolabs) and sequenced using the Nova SP 2x50bp protocol. ChIP-seq NEB Ultra protocol