GSM1423725: MCF10A_HC20_MYC-ER_ChIP-Seq; Homo sapiens; ChIP-Seq
Cells were cross-linked with 1% formaldehyde. After a hypotonic lysis step, nuclei were resuspended in sonication buffer containing 0.1% SDS and 0.1% Na-deoxycholate and sonication was performed in a Branson sonifier. Anti-ER-a antibody (HC20; Santa Cruz, sc-543) was coupled to Protein A-Dynabeads (Invitrogen). Bound chromatin was eluted with 1% SDS, and after decrosslinking, DNA was purified with phenol-chloroform. ChIP DNA was end-repaired, A-tailed and ligated to Illumina adaptors using the NEBNext ChIP-Seq Sample Preparation Kit. After size selection on an agarose gel and purification with a Qiagen gel extraction kit, PCR-amplification was performed. Library quality was tested with an Experion DNA Chip (BioRad) and quantified using a picogreen assay.