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SRX15212798: GSM6124010: GM12878_FC1; Homo sapiens; OTHER
1 OXFORD_NANOPORE (PromethION) run: 15M spots, 69.9G bases, 59.2Gb downloads

External Id: GSM6124010_r1
Submitted by: State Key Laboratory of Ophthalmology, Clinical Research Center for Ocular Disease, Zhongshan Ophthalmic Centre, Sun Yat-sen University
Study: in situ HiPore-C reveals higher-order 3D genome folding principles
show Abstracthide Abstract
Eukaryotic genomes are folded into three-dimensional (3D) hierarchical architectures consisting of multi-way chromatin contacts that are involved in complex gene regulation. High-throughput chromosome conformation capture (Hi-C), chromatin interaction analysis by paired-end tag sequencing (ChIA-PET) , and methods similar to these two rely on pairwise ligation of fragments in proximity in a population of cells, thus fail to reveal multi-way chromatin interactions in single allele at single fragment resolution. To explore higher-order chromatin interaction genome-widely, a straightforward strategy is integrating multi-fragment ligates preparation with third-generation sequencing technology. To achieve this purpose, we developed a protocol of in situ high throughput multi-way contact long read Pore-C sequencing (in situ HiPore-C) We performed genome-wide multi-way contact profiling analysis using data obtained from in situ HiPore-C sequencing of GM12878 and K562 cell lines. Overall design: in situ HiPore-C sequencing for GM12878 and K562 cell lines.
Sample: GM12878_FC1
SAMN28161071 • SRS12950403 • All experiments • All runs
Organism: Homo sapiens
Library:
Name: GSM6124010
Instrument: PromethION
Strategy: OTHER
Source: GENOMIC
Selection: other
Layout: SINGLE
Construction protocol: 15 million GM12878 or K562 cells were spun down and resuspended in 10 ml fresh medium. Cells were fixed with 1% formaldehyde and incubating for 10 minutes at room temperature (RT). After quenching, cell were lysed and cell nuclei were collected, then chromatin was digested with DpnII restriction enzyme and ligated with T4 DNA ligase, and recrosslinking reversal was performed with Proteinase K or Pronase. Finally DNA was purified using Phenol-Chloroform Isoamyl alcohol method. A total amount of 3-4ug purified DNA per sample was used as input material for the ONT library preparations. After the sample was qualified, size select of ligation DNA > 3kb were performed using the PippinHT system(Sage Science, USA). Next, the ends of DNA were repaired, and A-ligation reaction were conducted with NEBNext Ultra II End Repair/dA-tailing Kit(Cat# E7546). The adapter in the SQK-LSK109(Oxford Nanopore Technologies, UK) was used for further ligation reaction and DNA library was measured by Qubit 4.0 Fluorometer(Invitrogen, USA).
Runs: 1 run, 15M spots, 69.9G bases, 59.2Gb
Run# of Spots# of BasesSizePublished
SRR1914604814,973,83169.9G59.2Gb2023-01-20

ID:
21691122

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