GSM8678817: ESC, CEBPa induced, 24h, sorted SSEA1-, clone 1, input; Mus musculus; ChIP-Seq
For chromatin immunoprecipitations, 10 million cells were collected and crosslinked in 1% formaldehyde under rotation for 10 min, followed by glycine-mediated quenching. After the centrifuge, the cell pellet was re-suspended in SDS lysis buffer for 15 min and sonicated for 18 cycles of 30 sec on, and 30 sec off using the Bioruptor Pico (Diagenode). The supernatant was incubated with protein A magnetic beads (10002D, Invitrogen) pre-coupled with CEBPa antibody (8718, Cell Signaling) at 4℃ overnight. The next day, DNA was collected after sequentially washing and elution, followed by Proteinase K treatment and de-crosslinking. Libraries were prepared using the DNA Library Prep for Illumina kit (E7370, NEB) and amplified by Multiplex Oligos for Illumina kit (E6440, E6442, E6444, E6446, NEB). Libraries were sequenced on the NextSeq2000 instrument (Illumina), obtaining at least 40 million reads per sample.