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SRX19312009: GSM7032272: tandem601-OLTN; synthetic construct; MNase-Seq
1 ILLUMINA (Illumina MiSeq) run: 2.3M spots, 230.3M bases, 100.6Mb downloads

External Id: GSM7032272_r1
Submitted by: Division of Transcriptomics, Medical Institute of Bioregulation
Study: Analysis of the overlapping tri-nucleosome association with genomic and synthetic DNA
show Abstracthide Abstract
Nucleosome consists of a histone octamer wrapped by 145 bps of DNA and serves as a fundamental unit of chromatin. The overlapping di-nucleosome (OLDN) is found as another chromatin unit, in which 250 bps of DNA continuously wraps around the intimately associated histone octamer-hexamer complex. In the present study, our genome-wide analysis found a higher nucleosome stacking structure, overlapping tri-nucleosome (OLTN), in which about 350 bps of DNA wrap around the histone octamer-hexamer-hexamer complex. We quantified the length of DNA associated with the 20-mer histone core, and also we found that the OLTN localizes downstream of the transcription start sites as well as the OLDN does. The results suggest that nucleosome stacking structures may be formed for gene regulation. Overall design: MNase-seq of presumable fragments of OLTN, OLDN, and nucleosome using HeLa cell genomic DNA and synthetic DNA
Sample: tandem601-OLTN
SAMN33208971 • SRS16711049 • All experiments • All runs
Library:
Name: GSM7032272
Instrument: Illumina MiSeq
Strategy: MNase-Seq
Source: OTHER
Selection: MNase
Layout: PAIRED
Construction protocol: The HeLa cells were suspended in the washing solution containing 15 mM Tris-HCl (pH 8.0), 15 mM NaCl, 60 mM KCl, 300 mM sucrose, and a proteinase inhibitor (Roche). The detergent buffer containing 15 mM Tris-HCl (pH 8.0), 15 mM NaCl, 60 mM KCl, 300 mM sucrose, proteinase inhibitor (Roche), and 1% NP-40 was then added to the cell suspension, and the mixture was rotated at 4 ºC for 10 minutes. The HeLa nuclei were collected by centrifugation and were resuspended in the washing solution. A large amount of MNase (3 units/µL) and CaCl2 (2 mM) were added to the HeLa nuclei suspension. The MNase treatment was performed under 37 ºC for 30 minutes, and was terminated by EDTA (pH 8.0) addition. The MNase fragments were precipitated by ethanol, and were electrophoresed by 4% agarose gel. Libraries were prepared using ThruPLEX DNA-Seq Kit (Clontech, #R400674).
Runs: 1 run, 2.3M spots, 230.3M bases, 100.6Mb
Run# of Spots# of BasesSizePublished
SRR233709812,257,426230.3M100.6Mb2024-01-17

ID:
26558086

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