H3K27ac_WT2
Mus musculus
Protocols: Mouse hearts are perfused in a Langendorff apparatus and digested via liberase solution (Roche), the cells are going through different Ca2+ containing solutions. Cells are subcequently plated in a cellculture dish. The cells get fixated via incubation of 1% formaldehyde for 10 min at RT. The reaction is quenced using Glycin at a final concentration of 250 mM. Finally, the cells get collected and centrifuged to a cell pellet. The cells are lysed with the following buffers: Buffer1: 50mM Hepes‐KOH pH7.5, 140 mM NaCl, 1mM EDTA, 0.2ml 10% glycerol, 0.5% NP‐40, 0.25% Triton, X‐100 Buffer2: 200mM NaCl, 1mM EDTA (pH 8), 0.5mM EGTA (pH 8), 10mM Tris (pH 8) Buffer3: 50mM Tris (pH 8), 0,1% SDS, 0,95 % NP40, 0,1% Na-deoxycholate, 10mM EDTA, 150mM NaCl. The chromatin is fragmented via bioruptor sonification (Diagenode) using 4x10 cycles of sonification. The dna is precipitated using phenol/chloroform/isoamylalcohol and PhaseLock Tubes (5prime). Libraries are prepared according to the manufacture's protocol (NEBNext® ChIP-Seq Library Prep Master Mix Set for Illumina)
ENA-FIRST-PUBLIC
2019-01-31T17:02:07Z
ENA-LAST-UPDATE
2018-10-17T11:46:19Z
External Id
SAMEA4979766
INSDC center name
Heidelberg University Hospital, Department of Cardiology
INSDC first public
2019-01-31T17:02:07Z
INSDC last update
2018-10-17T11:46:19Z
INSDC status
public
Submitter Id
E-MTAB-7319:H3K27ac_WT2
broker name
ArrayExpress
cell type
cardiomyocyte
common name
house mouse
developmental stage
adult
genotype
wild type genotype
organism part
heart
replicate
2
sample name
E-MTAB-7319:H3K27ac_WT2
scientific_name
Mus musculus
sex
male