characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2023-05-11T14:01:52Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2023-05-11T14:01:52Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA111379527
characteristics.characteristic: External Id
characteristics.text: Universite Paris-Saclay, CEA, CNRS, Institute for Integrative Biology of the Cell (I2BC), 91198 Gif-sur-Yvette cedex, France
characteristics.characteristic: INSDC center name
characteristics.text: 2023-05-11T14:01:52Z
characteristics.characteristic: INSDC first public
characteristics.text: 2023-05-11T14:01:52Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS13476510
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-12198:MNase_input_Rsc8-Myc_med17-140
characteristics.characteristic: Submitter Id
characteristics.text: ArrayExpress
characteristics.characteristic: broker name
characteristics.text: Protocols: Following a shift at 37°C (duration depends on the mutant), after reaching exponential phase at 25°C in YPD, cells were cross-linked with 1% formaldehyde during 10 minutes. Cells were grown exponentially to 0.6 OD600 and cross-linked with 1% formaldehyde for 10 min. They were then washed with ST buffer (10 mM Tris-Cl pH 7.5, 100 mM NaCl) and frozen. Cells were lysed by bead-beating in FA lysis buffer (50 mM Hepes-KOH pH 8, 150 mM NaCl, 0.5 mM EDTA, 1% Triton, 0.1% sodium deoxycholate) supplemented with PMSF for 30 minutes at 4°C. Chromatin was recovered after a centrifuge step (16,000 rcf, 10 min, 4°C) in MNase buffer (20 mM Tris-HCl pH 7.5, 0.34 M sucrose, 15 mM KCl, 60 mM NaCl, 3 mM CaCl2) and was subjected to mild MNase (NEB, M0247S) treatment (20 min, 37°C on thermomixer). Reaction was stopped by addition of 10 mM EDTA on ice. Digested chromatin was recovered after centrifugation (16,000 rcf, 10 min, 4°C), and the remaining pellet was mildly sonicated (20” ON + 40” OFF, 4 cycles, mild), after a centrifugation step under similar conditions the chromatin samples were recovered at -80°C. The sizes of the DNA fragments have been checked by migrating the samples on both hand-made agarose and BioAanalyzer (Agilent) after pronase treatment. DNA was extracted with PCR purification kit (Qiagen). DNA was extracted using the Auto iPure V2 kit from Diagenode, following the manufacturer recommendations. Library construction was done according to the takara SMARTER thruplex (takara bio) protocol, following manufacturer's recommendation. Libraries were sized with AMPure beads (Beckman-coulter) to a final size of 325bp.
characteristics.characteristic: description
characteristics.text: MATa ura3-52 his3-∆200 ade2-101uaa trp1-∆63 lys2-801uag leu2-∆1 ∆med17::kan::ADE2 med17-140:CEN:TRP1 RSC8:13MYC:KANMX
characteristics.characteristic: genotype
characteristics.text: Shift at 37°C during 45min
characteristics.characteristic: growth condition
characteristics.text: RPM
characteristics.characteristic: normalization
characteristics.text: Saccharomyces cerevisiae S288C
characteristics.characteristic: organism
characteristics.text: Saccharomyces cerevisiae S288C
characteristics.characteristic: organism
characteristics.text: Saccharomyces cerevisiae S288C
characteristics.characteristic: scientific_name
characteristics.text: MNase_input_Rsc8-Myc_med17-140
characteristics.characteristic: title