GSM2113333: RNAPII 06hpi DMSO; Homo sapiens; ChIP-Seq
Crosslinking was performed for 10 minutes. For sonication, we used a refrigerated Bioruptor (Diagenode), which we optimized to generate DNA fragments of approximately 200– 1,000 base pair (bp). Lysates were pre-cleared for 3 hours using the appropriate isotype-matched control antibody (rabbit IgG; Cell Signaling) or anti-mouse IgG (Cell Signaling). The anti-RNA polymerase II (RNAPII) (clone 8WG16; Covance/BioLegend) antibodies were coupled with magnetic paramagnetic beads (Dynabeads® M-280 Sheep anti- 21 Mouse IgG; ThermoFisher Scientific) bound to anti–mouse IgG or anti–rabbit IgG for 6 hours. Antibody-bound beads and chromatin were then immune-precipitated overnight at 4 °C with rotation. After washing, reverse crosslinking was carried out overnight at 65°C. After digestion with RNase and proteinase K (Roche), DNA was isolated with a MinElute kit (Qiagen) and used for downstream applications.
GEO Accession
GSM2113333