L36_H3K27ac_rep_2
Homo sapiens
Protocols: Briefly, cells were crosslinked with 1% formaldehyde for 20 minutes and quenched by Glycine (125mM final concentration). Upon cell scraping, nuclear pellets were prepared in and washed with the nuclear preparation buffer (150mM NaCl, 20 mM EDTA, 50 mM Tris-HCl (pH 7.5), 0.5% v/v NP-40, 1% v/v Triton-X-100, 20 mM NaF). Then samples were sonicated in sonication buffer (150mM NaCl, 20 mM EDTA, 50 mM Tris-HCl (pH 8), 1% v/v NP-40, 0.5% v/v Sodium deoxycholate, 20 mM NaF, 0.1% SDS) for 30 cycles (in L3.6pl) and 25 cycles (in BxPC-3 and Panc-1) using a Bioruptor Pico (Diagenode) and a cycle setting of 30s on and 30s off. Consequently, samples were precleared by incubation with 50% slurry of Sepharose 4B (GE Healthcare) and incubated with antibody overnight. Antibodies included 1µg of p63 (4A4) (sc-8431, Santa-Cruz), 1µg of H3K27ac (196-050, Diagenode), 2µg of BRD4 (C15410337, Diagenode), and 1µg of Rabbit IgG (C15410206, Diagenode). Sepharose beads with Protein A (for rabbit antibodies) or Protein G (for mouse antibodies, 4A4 p63) were added to samples and incubated for 2 hours followed by washes, de-crosslinking, and DNA extraction. Extraction of DNA was performed using Roti(R)-phenol/chloroform/Isoamylalcohol (Roth) extraction followed by precipitation in 100% Ethanol and washes with 70% Ethanol. Libraries for DNA from ChIP were made using the Microplex Library Preparation kit v2 (Diagenode) according to the manufacturer's instructions.
ENA-FIRST-PUBLIC
2018-12-18T17:02:52Z
ENA-LAST-UPDATE
2018-07-20T14:51:18Z
External Id
SAMEA4799816
INSDC center name
Section of Tumor Epigenetics, Department of General, Visceral and Pediatric Surgery, University Medical Center Gottingen, 37075 Gottingen, Germany
INSDC first public
2018-12-18T17:02:52Z
INSDC last update
2018-07-20T14:51:18Z
INSDC status
public
Submitter Id
E-MTAB-7034:L36_H3K27ac_rep_2
broker name
ArrayExpress
cell line
L3.6pl
cell type
epithelial cell
common name
human
disease
pancreatic adenosquamous carcinoma
metastatic site
celiac lymph node
organism part
pancreas
sample name
E-MTAB-7034:L36_H3K27ac_rep_2
scientific_name
Homo sapiens