title: Input_ChIPseq_replicate1
organism: Homo sapiens
description: 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.
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2020-07-22T08:04:40Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2019-08-19T11:14:55Z
attributes.tag: External Id
attributes.value: SAMEA5871629
attributes.tag: INSDC center name
attributes.value: Department of Experimental Medicine 1, Nikolaus-Fiebiger-Center for Molecular Medicine, Friedrich-Alexander-University Erlangen-Nurnberg, Erlangen, Germany
attributes.tag: INSDC first public
attributes.value: 2020-07-22T08:04:40Z
attributes.tag: INSDC last update
attributes.value: 2019-08-19T11:14:55Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-8258:Input_ChIPseq_replicate1
attributes.tag: age
attributes.value: 51
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell line
attributes.value: MDAMB231
attributes.tag: cell type
attributes.value: mammary epithelial cell
attributes.tag: common name
attributes.value: human
attributes.tag: developmental stage
attributes.value: adult
attributes.tag: disease
attributes.value: breast adenocarcinoma
attributes.tag: genotype
attributes.value: wild type genotype
attributes.tag: organism part
attributes.value: mammary gland
attributes.tag: sample name
attributes.value: E-MTAB-8258:Input_ChIPseq_replicate1
attributes.tag: scientific_name
attributes.value: Homo sapiens