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Sample GSM3223248 Query DataSets for GSM3223248
Status Public on Jul 26, 2019
Title ATAC.d5FS.r1
Sample type SRA
 
Source name endoderm cells
Organism Mus musculus
Characteristics cell line: double knock-in (DKI) mouse ES cell line carrying Foxa2-Venus and Sox17-Cherry gene fusions
genotype/variation: Foxa2-Venus heterozygous;Sox17-Cherry homozygous
facs sorting markers: Foxa2-Venus-pos/Sox17-Cherry-pos
time point: d5
antibody: none
Growth protocol The cells were cultured for few consecutive passages on gelatine and ES medium. On the day of differentiation, mESCs were seeded on 10 cm gelatine coated dishes directly in endoderm differentiation medium (EDM) supplemented with 1 ng/ml of murine Wnt3a and 10 ng/ml of Activin A. Freshly prepared EDM supplemented with Wnt3a and Activin A was added every day. Cells were collected on day0, day3 and day5 for FACS isolation.
Extracted molecule genomic DNA
Extraction protocol ATAC-seq was done as previously described (Buenrostro et al., 2013). Briefly, 50000 FACS sorted cells were washed in 1xPBS, re-suspended in 50 ul of lysis buffer (10 mM Tris pH 7.4, 10 mM NaCl, 3 mM MgCl2, 0.1% NP40,) and spun at 500g for 10 min at 4 °C to collect nuclei. Nuclei were subsequently re-suspended in 50 µl Transposase reaction containing 25µL 2x tagmentation buffer, 22.5 µl water, 2.5 µl Tn5 Transposase (Illumina Nextera DNA Library Preparation Kit, cat. FC-121-1030). Reactions were incubated for 30 min at 37° C in a thermomixer shaking at 300 rpm and DNA purified using Qiagen PCR clean-up MinElute kit (Qiagen).
The transposed DNA was subsequently amplified in 50µl reactions with custom primers as described (Buenrostro et al., 2013). After 4 cycles, libraries were then monitored with qPCR: 5 µl PCR sample in a 15 µl reaction with the same primers. qPCR output was monitored for the ΔRN; 0.25 ΔRN cycle number was used to estimate the number of additional cycles of the PCR reaction needed for the remaining PCR samples. Amplified libraries were purified with the Qiagen PCR clean-up MinElute kit (Qiagen) and size selected for fragments less than 600 bp using the Agencourt AMPure XP beads (Beckman Coulter). Libraries were quality controlled by Qubit and Agilent DNA Bioanalyzer analysis.
 
Library strategy ATAC-seq
Library source genomic
Library selection other
Instrument model Illumina HiSeq 1500
 
Data processing Reads were mapped to the mouse genome (mm10) using bowtie (Langmead et al., 2009).
Genome_build: mm10 (GRCm38)
Supplementary_files_format_and_content: bigWig files were generated from Homer tag directories using makeBigWig.pl.
 
Submission date Jun 25, 2018
Last update date Jul 26, 2019
Contact name Filippo M. Cernilogar
E-mail(s) filippo.cernilogar@med.uni-muenchen.de
Organization name LMU Munich
Department Biomedical Center
Street address Grosshaderner Strasse 9
City Planegg-Martinsried
ZIP/Postal code 82152
Country Germany
 
Platform ID GPL18480
Series (2)
GSE116255 Pre-marked chromatin and transcription factor co-binding shape the pioneering activity of Foxa2 [ATAC-seq]
GSE116262 Pre-marked chromatin and transcription factor co-binding shape the pioneering activity of Foxa2
Relations
BioSample SAMN09486860
SRA SRX4298267

Supplementary file Size Download File type/resource
GSM3223248_ATAC.d5FS.r1_m1.ucsc.bigWig 242.2 Mb (ftp)(http) BIGWIG
SRA Run SelectorHelp
Raw data are available in SRA
Processed data provided as supplementary file

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