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Sample GSM3756149 Query DataSets for GSM3756149
Status Public on Dec 11, 2019
Title Hi-C MCF7 T0 Replicate 1
Sample type SRA
 
Source name MCF7
Organism Homo sapiens
Characteristics cell line: MCF7
tissue: breast cancer
resistant phynotype: responsive
Growth protocol MCF7 breast cancer cells were kindly given to our laboratory by Dr Liz Caldon (Garvan Institute, Australia). MCF7 cells were maintained in RPMI-1640 based medium containing 5% (v/v) fetal calf serum (FCS). All cell lines were authenticated by short-tandem repeat (STR) profiling (Cell Bank, Australia) and cultured for less than 6 months after authentication.
Extracted molecule genomic DNA
Extraction protocol Cells were cross-linked with formaldehyde, lysed and the intact nuclei was permeabilized. DNA was then cut with NcolI restriction enzyme, the overhangs filled in incorporating a biotinylated base and the free ends were then ligated together in situ. Crosslinks were reversed, the DNA was sheared to 300-500bp and the biotinylated ligation junctions were pulled down with streptavidin beads.
Custom library construction protocol was followed. Briefly, DNA was end-repaired and dA-tailing using the NEBNext DNA Master Mixes kit and the ligation of universal adapter was performed on Streptavidin-bound DNA using the NEBNext Ultra DNA Library Prep kit. After adapter ligation DNA was PCR amplified with indexed Illumina primers for 8-12 cycles and library fragments of at least 200bp were purified using SPRI beads. The purified DNA was captured on an Illumina flow cell for cluster generation. Libraries were sequenced on the HiSeq2500 following the manufacturer's protocols.
 
Library strategy Hi-C
Library source genomic
Library selection other
Instrument model Illumina NextSeq 500
 
Data processing Raw sequence data was mapped and processed using HiC-Pro v2.7.7 (Servant et al., 2015). The paired end reads were aligned separately using bowtie2 against the hg38 human build. Contact matrices were constructed at multiple resolutions and normalized suing ICE normalization algorithm (Imakaev et al. 2012).
Juicebox hic files were prepared using hicpro2juicebox script (Servant et al., 2015).
Genome_build: GRCh38/hg38
Supplementary_files_format_and_content: allValidPairs files contain all valid interactions identified at multiple resolutions (10kb, 20kb, 40kb, 150kb, 500kb and 1Mb) using HiC-Pro pipeline. The validPairs are stored using a tab-delimited text format as described in (Servant et al., 2015). hic files contain compressed contact matrices at all resolutions. hic format can also be used to visualise Hi-C data in Juicebox (Durand et al., 2016; Robinson et al., 2018) or WashU Epigenome Browser (Zhou et al., 2013).
 
Submission date May 08, 2019
Last update date Dec 13, 2019
Contact name Joanna Achinger-Kawecka
E-mail(s) j.achinger@garvan.org.au
Organization name Garvan Institute of Medical Research
Lab Epigenetics Research Laboratory
Street address 384 Victoria Street
City Darlinghurst
State/province NSW
ZIP/Postal code 2010
Country Australia
 
Platform ID GPL18573
Series (1)
GSE130916 Deregulation of the three-dimensional epigenome in endocrine resistant breast cancer
Relations
BioSample SAMN11605338
SRA SRX5807390

Supplementary file Size Download File type/resource
GSM3756149_20190418_Hi-C_MCF7Caldon_T0_a1_allValidPairs.hic 459.3 Mb (ftp)(http) HIC
GSM3756149_20190418_Hi-C_MCF7Caldon_T0_a1_allValidPairs.txt.gz 644.5 Mb (ftp)(http) TXT
SRA Run SelectorHelp
Raw data are available in SRA
Processed data provided as supplementary file

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