GSM4678343: MCF10A siNT  RNAPII ChIP-seq; Homo sapiens; ChIP-Seq
MCF10A cells were fixed in 10% fixing solution (11% formaldehyde, 0.1 M NaCl, 1 mM EDTA (pH 8.0), and 50 mM HEPES (pH 7.9)) for 15 minutes at room temperature. Cross-linking was stopped with 5% v/v 2.5 M glycine at room temperature for 5 minutes. Cells were washed once in PBS-Igepal (0.5%). Cells were washed again in PBS-Igepal (0.5%) with 1 mM PMSF. The cells were centrifuged at 800 x g, the supernatant removed, and the cell pellet was flash frozen on dry ice. Further steps of the TranscriptionPath RNAPII ChIP-seq protocol were performed by Active Motif. Chromatin immunoprecipitation was completed using an RNAPII antibody (Active Motif, Clone H4B, Cat. No. 39097). Sequencing was performed using the Illumina NextSeq 500 sequencing system with 75 nt sequence reads.
GEO Accession
GSM4678343