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Sample GSM1621288 Query DataSets for GSM1621288
Status Public on May 08, 2015
Title mm1036_WT_mm9
Sample type SRA
 
Source name distal forelimb E11.5
Organism Mus musculus
Characteristics strain: 129/Sv x C57BL/6
tissue: distal forelimb
age: E11.5
genotype: wild type
1st restriction enzymes: BglII
2nd restriction enzymes: Csp6I
viewpoint: mm1036
Treatment protocol Distal forelimbs were microdissected and dissociated using a Trypsin solution (0.05%).
Growth protocol E11.5 mouse embryos were collected in ice cold 10% FCS in PBS
Extracted molecule genomic DNA
Extraction protocol 4C-seq libraries were generated from microdissected tissues or cells as described previously (van de Werken et al., 2012). BglII or HindIII (6-bp cutters) were used as primary restriction enzymes. Csp6I or DpnII were used as secondary restriction enzymes. For each viewpoint, a total of 1.6 mg of each library was amplified in 8 or 16 PCR reactions, depending on PCR efficiencies. Samples were multiplexed and sequenced with Ilumina Hi-Seq technology. The following primers were used: Wnt6_promoter_forward: CTACACGACGCTCTTCCGATCTCCACAGATGCCTACATCAG, Wnt6_promoter_reverse: CAGACGTGTGCTCTTCCGATCTCAGTTGGACAGCCTTCTACC, Ihh promoter_forward: CTACACGACGCTCTTCCGATCTCTTGCTCGGGATTCAGAT, Ihh_promoter_reverse: CAGACGTGTGCTCTTCCGATCTGCATGTCTAGCATGGTGGTA, mm1036_forward: CTACACGACGCTCTTCCGATCTACCTTATCTTGGCCTTCAA, mm1036_reverse: CAGACGTGTGCTCTTCCGATCTCCATAACCCAGACTGAGATG, Epha4_promoter_forward: CTACACGACGCTCTTCCGATCTTGGCTTTTACCGAAAATAAA, Epha4_promoter_reverse: CAGACGTGTGCTCTTCCGATCTTCGTAGTCTAGGTGGAGGAA Pax3_promoter_forward: CTACACGACGCTCTTCCGATCTTTCTATCTCCCCATGACACC, Pax3_promoter_reverse: CAGACGTGTGCTCTTCCGATCTTCCTCCAAACGTGCTCTG
Samples were multiplexed and sequenced with Ilumina Hi-Seq technology according to the manufacturer´s protocol.
 
Library strategy OTHER
Library source genomic
Library selection other
Instrument model Illumina HiSeq 1000
 
Data processing Library strategy: 4C-Seq
Reads were mapped by Novoalign (Novoalign V2.07; www.novocraft.com) to the reference sequences NCBI37/mm9.
To visualize the data, we created files in bedGraph track format for the readcounts of each fragment or in a specified window of fragments.
The viewpoint, adjacent undigested fragment and fragments 10 kb up and downstream were removed.
4C-seq contacts were analyzed in the mouse region chr1:73000000-79000000. A range of 5 fragments was used to normalize the data per million reads (RPM) over a sliding window and display the continuous-valued data in the figures. Log2 ratios were calculated by dividing fragment reads between different samples.
Genome_build: mm9
Supplementary_files_format_and_content: Smoothed/normalized data (bedGraph).
 
Submission date Feb 27, 2015
Last update date May 15, 2019
Contact name Darío G Lupiáñez
E-mail(s) lupianez@molgen.mpg.de
Organization name Max Planck Institute for Molecular Genetics
Street address Ihnestrasse 73
City Berlin
ZIP/Postal code 14195
Country Germany
 
Platform ID GPL15103
Series (2)
GSE66379 Disruptions of Topological Chromatin Domains Causes Pathogenic Rewiring of Gene-Enhancer Interactions [4C-Seq-mouse]
GSE66383 Disruptions of Topological Chromatin Domains Causes Pathogenic Rewiring of Gene-Enhancer Interactions
Relations
BioSample SAMN03379905
SRA SRX893606

Supplementary file Size Download File type/resource
GSM1621288_mm1036_WT_mm9.bedgraph.gz 5.4 Mb (ftp)(http) BEDGRAPH
SRA Run SelectorHelp
Raw data are available in SRA
Processed data provided as supplementary file

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