characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2022-03-28T12:20:30Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2022-03-28T12:20:30Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA12595084
characteristics.characteristic: External Id
characteristics.text: High Definition Disease Modelling Lab, Stem Cell and Organoid Epigenetics, IEO, European Institute of Oncology, IRCCS, Milan, Italy. Human Technopole.
characteristics.characteristic: INSDC center name
characteristics.text: 2022-03-28T12:20:30Z
characteristics.characteristic: INSDC first public
characteristics.text: 2022-03-28T12:20:30Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS10201562
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-11363:11_10DTAMO_H3K27me3
characteristics.characteristic: Submitter Id
characteristics.text: 18
characteristics.unit: day
characteristics.characteristic: age
characteristics.text: ArrayExpress
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0009737
characteristics.characteristic: broker name
characteristics.text: ESC derived cell line
characteristics.characteristic: cell line
characteristics.text: differentiating neuron
characteristics.characteristic: cell type
characteristics.text: house mouse
characteristics.characteristic: common name
characteristics.text: Protocols: Cells at the various time-points (72 hours or 10 days after Cre induction) were cross-linked in 1% formaldehyde for 10 min and the crossliking was stopped by incubating in 0.125 M glycine for 5 min. After washing cells were harvested in SDS buffer (0.5% SDS, 50mM Tris-Cl pH 8.1, 100mM NaCl, 5mM EDTA pH 8, 0.02% NaN3). Prior to sonication, the lysate was diluted 2:1with dilution buffer (5% Triton X-100, 100mM Tris-Cl pH 8.6, 100mM NaCl, 5mM EDTA pH 8.5, 0.02% NaN3). Sonication was performed with Covaris sonicator generating DNA fragments with a bulk size around 250bp. Chromatin was quantified with Lowry method. After sonication, 600μg of chromatin and 12μg of anti-H3K27me3 (#9733BF Cell Signalling) were used for immunoprecipitation overnight at 4°C and recovered the following day using Dynabeads Protein G (Life Technologies). 1% of chromatin prior to immunoprecipitation was used as input. Immunoprecipitate was washed three times in low salt and one time in high salt wash buffer (1% Triton-X, 150mM or 500mM NaCl, 20mM Tris-Cl pH 8.0, 0.1% SDS, 2mM EDTA pH 8.0). One additional wash was performed in TE. Elution and crosslink reverse was performed under standard condition. DNA was then purified using PCR purification kit (QIAGEN). Libraries were prepared by the sequencing facility at IEO according to the protocol described by Blecher-Gonen and colleagues (PMID: 23429716)
characteristics.characteristic: description
characteristics.text: normal
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/PATO_0000461
characteristics.characteristic: disease
characteristics.text: Ezh2 fl/fl
characteristics.characteristic: genotype
characteristics.text: Mus musculus
characteristics.characteristic: organism
characteristics.text: Mus musculus
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_10090
characteristics.characteristic: organism
characteristics.text: embryonic stem cell
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/CL_0002322
characteristics.characteristic: progenitor cell type
characteristics.text: Line2
characteristics.characteristic: replicate
characteristics.text: 8 days + 10 days after Cre induction
characteristics.characteristic: sampling time point
characteristics.text: Mus musculus
characteristics.characteristic: scientific_name
characteristics.text: 4-hydroxytamoxifen
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/CHEBI_44616
characteristics.characteristic: stimulus
characteristics.text: 11_10DTAMO_H3K27me3
characteristics.characteristic: title