GSM1540467: Input con; Homo sapiens; ChIP-Seq
primary keratinocytes cultured in eight 30-cm plates were scraped off the dish with PBS at about 60-80% confluence, and cross-linked with sequentially with 1% formaldehyde for 8 mins, then treated with 1.25M glycine for 5 min to stop crosslink. Cells were pelleted by centrifuging 5 min at 1500 rpm, and washed twice with PBS (1mM PMSF). Cell nuclei was isolated following cell lysis with ChIP lysis buffer and resuspended with complete RIPA buffer (Santa Cruz Biotechnology, Santa CruZ, CA) for DNA fragmentation by sonication with Branson Sonifier. The fragmented DNA underwent IP with the antibody against JunB (Cell Signaling Technology) followed by purification with MinElute DNA kit (Qiagen). 10 ng of the purified DNA was used for library construction with - the NEBNext® DNA library preparation kit (New England BioLabs).
GEO Accession
GSM1540467