Input_ChIPseq_replicate2
Homo sapiens
Protocols: Chromatin was cross-linked by 1.5mM EGS for 30min followed by 1% formaldehyde for 10min at room temperature (RT). Crosslinking was stopped by adding glycine to a final concentration of 125mM for 5min at RT. Cells were lysed in NP40 lysis buffer containing protease inhibitors and disrupted by dounce homogenisation. MDA-MB-231 cells were purchased from the American Type Culture Collection (ATCC) and cultured under standard conditions in DMEM (Gibco) supplemented with 10 % fetal bovine serum (Gibco). Nulcei were lysed and sonicated with a Diagenode Bioruptor to a fragment size of 300 to 500bp. Chromatin was collected and precleared with protein A and G Dynabeads. Chromatin-immunoprecipitation was performed using an anti-ZEB1 antibody (Santa Cruz, SC-H102-X). Immuncomplexes were captured using Dnyabeads and DNA was extracted by QIAquick columns (MinElute). Several IPs were pooled for sequencing. Library preparation was performed with the NEBNext® Ultra DNA Library Prep Kit according to the manufacturer's instructions.
ENA-FIRST-PUBLIC
2020-07-22T08:04:40Z
ENA-LAST-UPDATE
2019-08-19T11:14:55Z
External Id
SAMEA5871630
INSDC center name
Department of Experimental Medicine 1, Nikolaus-Fiebiger-Center for Molecular Medicine, Friedrich-Alexander-University Erlangen-Nurnberg, Erlangen, Germany
INSDC first public
2020-07-22T08:04:40Z
INSDC last update
2019-08-19T11:14:55Z
INSDC status
public
Submitter Id
E-MTAB-8258:Input_ChIPseq_replicate2
age
51
broker name
ArrayExpress
cell line
MDAMB231
cell type
mammary epithelial cell
common name
human
developmental stage
adult
disease
breast adenocarcinoma
genotype
wild type genotype
organism part
mammary gland
sample name
E-MTAB-8258:Input_ChIPseq_replicate2
scientific_name
Homo sapiens