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Sample GSM5034633 Query DataSets for GSM5034633
Status Public on Jul 20, 2021
Title E12.5 inv2 proximal forelimb ATAC-seq
Sample type SRA
 
Source name proximal forelimb
Organism Mus musculus
Characteristics age: E12.5
strain: B6CBAF1
genotype: inv2
tissue: proximal forelimb
Treatment protocol No treatment
Growth protocol Crosses were made between heterozygous adults. Animals were checked for vaginal plugs on the next morning, establishing the timepoint 0.5 embryonic days. Embryos were collected 12 days later.
Extracted molecule genomic DNA
Extraction protocol The forelimbs of embryos were dissected and placed into PBS with 10% FCS and 8ul collagenase at 50mg/ml (Sigma C9697) at 37ยบ for approximately 5min. Cells were counted and 50,000 cells were isolated for processing with the Nextera Tn5 enzyme (Illumina FC-131-1096) as previously described (Buenrostro NatMeth 2013).
Tn5 treated DNA was amplified with Nextera Library primers using NEBNext library amplification master mix (NEB M0541).
 
Library strategy ATAC-seq
Library source genomic
Library selection other
Instrument model Illumina NextSeq 500
 
Data processing ATAC-seq reads processing was done on the Duboule local Galaxy server (Afgan et al., 2016). Reads were mapped to the mouse genome (mm10) using Bowtie2 (v2.3.4.1) (Langmead and Salzberg, 2012), (-I 0 -X 2000 --fr --dovetail --very-sensitive-local).
Reads with mapping quality below 30, mapping to mitochondria, or not properly paired were removed from the analysis. PCR duplicates were filtered using Picard (v1.56.0).
BAM file was converted to BED with bedtools version 2.27. Peak calling was done using MACS2 (v2.1.0.20151222) callpeak (--nomodel --shift 100 --extsize 200 --call-summits --keep-dup all).
The coverage was done using the center of the Tn5 insertion and extended on both sides by 20 bp (script available at https://github.com/lldelisle/scriptsForBoltEtAl2021)
Genome_build: mm10
Supplementary_files_format_and_content: bigwig: coverage centered on the Tn5 insertion (20bp extended both sides)
narrowPeak: narrowPeak from macs2
 
Submission date Jan 25, 2021
Last update date Jul 20, 2021
Contact name Lucille Lopez-Delisle
E-mail(s) lucille.delisle@epfl.ch
Organization name EPFL
Street address Station 19
City Lausanne
ZIP/Postal code 1015
Country Switzerland
 
Platform ID GPL19057
Series (2)
GSE165490 Mesomelic Dysplasias Associated With The HOXD Locus Are Caused By Regulatory Reallocations [ATAC-Seq]
GSE165495 Mesomelic Dysplasias Associated With The HOXD Locus Are Caused By Regulatory Reallocations
Relations
BioSample SAMN17531913
SRA SRX9934230

Supplementary file Size Download File type/resource
GSM5034633_E12.5_inv2_PFL_ATAC.bw 202.0 Mb (ftp)(http) BW
GSM5034633_E12.5_inv2_PFL_ATAC.narrowPeak.gz 2.8 Mb (ftp)(http) NARROWPEAK
SRA Run SelectorHelp
Raw data are available in SRA
Processed data provided as supplementary file

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