GSM1948651: S5p_K52o_EtOH; Homo sapiens; ChIP-Seq
Where indicated, MYC K52o-HA was induced with doxycycline (pInducer samples) or treated with EtOH as solvent control. Where indicated MG-132 (10 µM) or EtOH as a control was added to the cells 4 hr prior to fixation. Cells were crosslinked with 1% formaldehyde for 10min. Cells were lysed and after centrifugation nuclei were re-suspended in RIPA buffer. DNA was sonicated with a Branson sonifier to obtain DNA fragments at nucleosomal size. Chromatin was precipitated by incubation with specific antibodies pre-adsorbed to Protein A dynabeads beads overnight. After several washings chromatin was eluted with 1 % SDS and  crosslinking was reverted overnight. DNA was purified by Phenol/Chloroform extraction and enrichments were checked by qPCR. Libraries for ChIP-seq samples were contructed following manufactor's intructions using the NEBNext ChIP-Seq Library Prep Master Mix Set for Illumina (E6240). Briefly, ChIP DNA was end repaired,  A-tailed  and Illumina adaptors were ligated. DNA fragments of about 200 bps were cut out of a agarose gel and extracted with a Qiagen PCR purification column. Size-selected DNA was amplified with 18 PCR cycles.
GEO Accession
GSM1948651