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SRX23224030: GSM8017946: wild type Mre11-HA6 t5 WCE; Saccharomyces cerevisiae; Nakaseomyces glabratus; ChIP-Seq
1 ILLUMINA (Illumina HiSeq 2500) run: 5.3M spots, 791.9M bases, 278.4Mb downloads

External Id: GSM8017946_r1
Submitted by: Chromosome Biology, Max Perutz Labs, University of Vienna
Study: Physical interaction with Spo11 mediates the localisation of Mre11 to chromatin in meiosis and promotes its nuclease activity
show Abstracthide Abstract
Meiotic recombination is of central importance for the proper segregation of homologous chromosomes and is initiated by DNA double-strand breaks (DSB) that depend on Spo11 and Mre11. Calibrated chromatin immunoprecipitation (ChIP) for Mre11 shows that Spo11 promotes Mre11 recruitment to chromatin, independent of DSB formation. A C-terminal deletion mutant deficient in Spo11 interaction severely reduces the association of Mre11 with meiotic chromatin. Consistent with the reduction of Mre11 foci in this mutant, it strongly impedes DSB formation, leading to spore death. Overall design: Six ChIP-seq samples and six corresponding whole cell extract (WCE) samples were derived from Saccharomyces cerevisiae (S. cer.) SK1 cells with HA-tagged (or untagged) Mre11 grown in meiotic culture for 5 hrs ("t5") and mixed with Candida glabrata (C. gla., with Scc1-HA6) CBS138 cells for calibration and downstream quantiative comparison. Paired-end (75bp) fastq.gz files are provided, as well as calibrated processed files ("filled dpp" with chromosome, position, and depth information).
Sample: wild type Mre11-HA6 t5 WCE
SAMN39447973 • SRS20147240 • All experiments • All runs
Library:
Name: GSM8017946
Instrument: Illumina HiSeq 2500
Strategy: ChIP-Seq
Source: GENOMIC
Selection: ChIP
Layout: PAIRED
Construction protocol: Cells were opened by FastPrep-24™ 5G Instrument (MP Biomedicals) followed by sonication with Bioruptor® Pico instrument. After removing the cell debris, the supernatant was used for the ChIP with anti-HA antibody. To reverse crosslink, all the samples were incubated in 65°C overnight. DNA purification was conducted by regular phenol/chloroform/isoamyl alcohol extraction. Library preparation was performed using NEBNext Ultra II DNA Library Prep Kit according to the manufacturer´s protocol. Amplified library DNA was purified and subjected to 300 bp - 500 bp size selection using Agencourt AMPure XP beads (Beckman Coulter) and quantified by NanoDrop 3300 Fluorospectrometer and Agilent 2100 Bioanalyzer.
Runs: 1 run, 5.3M spots, 791.9M bases, 278.4Mb
Run# of Spots# of BasesSizePublished
SRR275546905,279,015791.9M278.4Mb2024-01-25

ID:
31380180

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