GSM2071223: Erα_Chipseq_Y537S_INPUT; Homo sapiens; ChIP-Seq
Chromatin immunoprecipitation (ChIP) was performed in MCF7_LUC and MCF7_Y537S cell lines. The cells were seeded at 10X10^6 cells/plate on 150cm dishes in DMEM lacking of phenol-red with 10% dextran-coated charcoal stripped FCS for 72 hours, then treated with ethanol/oestrogen (10nM) for 45 mins prior to fixation. Lysates were collected from sonicated nuclei extracts and subsequently were immuno-precipitated with Erα antibody (HC-20, santacruz). DNA was then extracted using phenol-chloroform and sodium chloride precipitation method. For Chip-seq, 10ng DNA from IP and input samples were used for library preparation. Libraries were prepared according to NEB (E7370S) multiplex library preparation protocol for DNA-seq preparation. Libraries were run on bioanalyzer for quality control. Finally, sequencing was performed on Ilumina Hiseq 2000 genome analyser (using 50b reads)