characteristics.text: 2014-01-21T17:01:05Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2018-03-08T16:12:57Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA2045033
characteristics.characteristic: External Id
characteristics.text: University of Cambridge
characteristics.characteristic: INSDC center name
characteristics.text: 2014-01-21T17:01:05Z
characteristics.characteristic: INSDC first public
characteristics.text: 2018-03-08T16:12:57Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS205785
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-1447:IgG Control
characteristics.characteristic: Submitter Id
characteristics.text: 2 year
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/UO_0000036
characteristics.characteristic: age
characteristics.text: Takehido Kobayashi
characteristics.characteristic: biosource provider
characteristics.text: frozen sample
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/OBI_0000922
characteristics.characteristic: biosource type
characteristics.text: ArrayExpress
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0009737
characteristics.characteristic: broker name
characteristics.text: baker's yeast
characteristics.characteristic: common name
characteristics.text: Protocols: Yeast cells were grown to mid-logarithmic phase before fixation Cells were fixed with 1% formaldehyde for 45 minutes at room temperature, washed with PBS and resuspended in 500ul of SDS lysis buffer (1% SDS, 10mM EDTA, 50mM Tris HCl (8.0), 0.5mM PMSF-EtOH, 0.8_g/ml pepstatin A, 0.6_g/ml leupeptin). Cells were lysed by bead-beating 4x 1min with 3min on ice in between. Cell debris was removed by centrifugation and lysates were sonified for 30min in a constantly water-cooled Bioruptor. Sonified lysates were centrifuged and the supernatant was diluted 1:10 into IP-buffer (0.01% SDS, 1.1%Triton-X-100, 1.2mM EDTA, 16.7MM Tris HCl (8.0), 167mM NaCl, 0.5mM PMSF, 0.8_g/ml Pepstatin A, 0.6_g/ml leupeptin). Antibodies were added (1ug each of IgG (ab37415), H2A (Active Motif 39235) and 1/500 dilution of H2AQ105me) over night. Next day, 30ul of a Protein A/G Dynabead mix was added and beads were incubated for 1h. Subsequently, beads were washed with 1ml each TSE150 (1% Triton-X-100, 0.1%SDS, 2mM EDTA, 20mM Tris HCl (8.0), 150mM NaCl), TSE500 (1% Triton-X-100, 0.1%SDS, 2mM EDTA, 20mM Tris HCl (8.0), 500mM NaCl), and LiCl-buffer (0.25M LiCl, 1% NP-40, 1% dioxycholate, 1mM EDTA, 10mM Tris HCl (pH8.0)). Finally, beads were wahed with 1ml TE and eluted with elution buffer (1% SDS, 0.1M NaHCO3) for 30min at room temperature. 500mM NaCL (final) was added and crosslinks were reversed at 95oC for 30min. Samples were treated with 50ug RNase/DNase free. Samples were purified with the Qiagen PCR purification kit following manufacturer's guidelines.
characteristics.characteristic: description
characteristics.text: Mata ade2-1 ura3-1 his3-11 trp1-1 leu2-3,112 can1-100 fob1D::LEU2 (20 rDNA copies)
characteristics.characteristic: genotype
characteristics.text: Saccharomyces cerevisiae
characteristics.characteristic: organism
characteristics.text: Saccharomyces cerevisiae
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_4932
characteristics.characteristic: organism
characteristics.text: Saccharomyces cerevisiae
characteristics.characteristic: scientific_name
characteristics.text: YSI1102
characteristics.characteristic: strain or line
characteristics.text: IgG Control
characteristics.characteristic: title