GSM3107931: siCTRL_Pol2 ChIPSeq; Homo sapiens; ChIP-Seq
Lysates were sonicated and the DNA  sheared to an average length of 300-500 bp. Genomic DNA (Input) was prepared by treating aliquots of  chromatin with RNase, proteinase K and heat for de-crosslinking, followed by ethanol precipitation. Pellets  were resuspended and the resulting DNA was quantified on a NanoDrop spectrophotometer. Extrapolation to  the original chromatin volume allowed quantitation of the total chromatin yield. An aliquot of chromatin (30 ug) was precleared with protein A (for Nrf2) or G (for Pol2) agarose beads  (Invitrogen). Genomic DNA regions of interest were isolated using 4 ug of target specific antibody.  Complexes were washed, eluted from the beads with SDS buffer, and subjected to RNase and proteinase K  treatment. Crosslinks were reversed by incubation overnight at 65 C, and ChIP DNA was purified by phenolchloroform  extraction and ethanol precipitation. Illumina sequencing libraries  were prepared from the ChIP and Input DNAs by the standard consecutive enzymatic steps of end-polishing,  dA-addition, and adaptor ligation. After a final PCR amplification step, the resulting DNA libraries were  quantified and sequenced on Illumina's NextSeq 500 (75 nt reads, single end).
GEO Accession
GSM3107931