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Pol2_Rsc8-Myc_444

Identifiers
BioSample: SAMEA111379532; SRA: ERS13476515
Organism
Saccharomyces cerevisiae S288C
cellular organisms; Eukaryota; Opisthokonta; Fungi; Dikarya; Ascomycota; saccharomyceta; Saccharomycotina; Saccharomycetes; Saccharomycetales; Saccharomycetaceae; Saccharomyces; Saccharomyces cerevisiae
Attributes
genotypeMATa ura3-52 his3-∆200 ade2-101uaa trp1-∆63 lys2-801uag leu2-∆1 ∆med17::kan::ADE2 med17-444:CEN:TRP1 RSC8:13MYC:KANMX
sample nameE-MTAB-12198:Pol2_Rsc8-Myc_444
ENA-CHECKLISTERC000011
ENA-FIRST-PUBLIC2023-05-11T14:01:52Z
ENA-LAST-UPDATE2023-05-11T14:01:52Z
External IdSAMEA111379532
INSDC center nameUniversite Paris-Saclay, CEA, CNRS, Institute for Integrative Biology of the Cell (I2BC), 91198 Gif-sur-Yvette cedex, France
INSDC first public2023-05-11T14:01:52Z
INSDC last update2023-05-11T14:01:52Z
INSDC statuspublic
Submitter IdE-MTAB-12198:Pol2_Rsc8-Myc_444
broker nameArrayExpress
growth conditionShift at 37°C during 45min
normalizationSpike-in
scientific_nameSaccharomyces cerevisiae S288C
Description

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.

BioProject
PRJEB56195 Functional interplay between Mediator and RSC chromatin remodeling complex in nucleosome-depleted region maintenance at promoters
Retrieve all samples from this project

Submission
EBI; 2023-05-19
Accession:
SAMEA111379532
ID:
35185229

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