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| Status |
Public on Oct 13, 2017 |
| Title |
Rao-2017-HIC028 |
| Sample type |
SRA |
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| Source name |
Colorectal carcinoma
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| Organism |
Homo sapiens |
| Characteristics |
cell line: HCT-116-RAD21-mAC
protocol: in situ Hi-C
treatment: 500uM auxin (360 min)
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| Treatment protocol |
Cells were treated with 500uM auxin as described in the characteristics above
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| Growth protocol |
Cell lines were cultured according to manufacturer's instructions
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| Extracted molecule |
genomic DNA |
| Extraction protocol |
Cells were crosslinked and then lysed with nuclei permeabilized but still intact. DNA was then restricted and the overhangs filled in incorporating a biotinylated base. Free ends were then ligated together in situ. Crosslinks were reversed, the DNA was sheared to 300-500bp and then biotinylated ligation junctions were recovered with streptavidin beads.
standard Illumina library construction protocol, Briefly, DNA was end-repaired using a combination of T4 DNA polymerase, E. coli DNA Pol I large fragment (Klenow polymerase) and T4 polynucleotide kinase. The blunt, phosphorylated ends were treated with Klenow fragment (32 to 52 exo minus) and dATP to yield a protruding 3- 'A' base for ligation of Illumina's adapters which have a single 'T' base overhang at the 3’ end. After adapter ligation DNA was PCR amplified with Illumina primers for 8-12 cycles and library fragments of 400-600 bp (insert plus adaptor and PCR primer sequences) were purified using SPRI beads. The purified DNA was captured on an Illumina flow cell for cluster generation. Libraries were sequenced on the HiSeq X Ten, Illumina NextSeq 500, or HiSeq 2500 following the manufacturer's protocols
Hi-C; Specific library strategies are indicated as additional columns in the SAMPLES section
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| Library strategy |
Hi-C |
| Library source |
genomic |
| Library selection |
other |
| Instrument model |
HiSeq X Ten |
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| Data processing |
The paired end reads were aligned separately using BWA against the b37 human build
PCR duplicates, low mapping quality and unligated reads were removed using Juicer (see Rao, Huntley, et al Cell 2014; Durand, Shamim, et al Cell Systems 2016)
Contact matrices were constructed at various resolutions and normalized using Juicer (see Rao, Huntley, et al Cell 2014; Durand, Shamim, et al Cell Systems 2016)
loops were annotated using HiCCUPS (see Rao, Huntley, et al Cell 2014; Durand, Shamim, et al Cell Systems 2016), domains were annotated using Arrowhead (see Rao, Huntley, et al Cell 2014; Durand, Shamim, et al Cell Systems 2016)
Genome_build: b37 (human)
Supplementary_files_format_and_content: .hic file (contains contact matrices at various resolutions in an easy to visualize and access format) (see Durand, Robinson, et al Cell Systems 2016)
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| Submission date |
Oct 12, 2017 |
| Last update date |
May 15, 2019 |
| Contact name |
Suhas Rao |
| E-mail(s) |
suhasrao@post.harvard.edu
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| Organization name |
Baylor College of Medicine
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| Department |
Molecular and Human Genetics
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| Lab |
The Center for Genome Architecture
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| Street address |
1 Baylor Plaza
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| City |
Houston |
| State/province |
TX |
| ZIP/Postal code |
77030 |
| Country |
USA |
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| Platform ID |
GPL20795 |
| Series (2) |
| GSE104333 |
Cohesin loss eliminates all loop domains [HiC] |
| GSE104334 |
Cohesin loss eliminates all loop domains |
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| Relations |
| BioSample |
SAMN07777209 |
| SRA |
SRX3276127 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSM2809559_Rao-2017-HIC028.hic |
364.8 Mb |
(ftp)(http) |
HIC |
| GSM2809559_Rao-2017-HIC028_30.hic |
343.0 Mb |
(ftp)(http) |
HIC |
SRA Run Selector |
| Raw data are available in SRA |
| Processed data provided as supplementary file |
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