Sample information curated by ChIP-Atlas

Antigen

Antigen Class
No description
Antigen
NA

Cell type

Cell type Class
Pluripotent stem cell
Cell type
ES cells
NA
NA

Attributes by original data submitter

Sample

ENA first public
2019-02-17
ENA last update
2018-07-16
ENA-CHECKLIST
ERC000011
External Id
SAMEA4794057
INSDC center alias
Philipps-University Marburg, Genomics Core Facility
INSDC center name
Philipps-University Marburg, Genomics Core Facility
INSDC first public
2019-02-17T17:02:49Z
INSDC last update
2018-07-16T17:07:27Z
INSDC status
public
Submitter Id
E-MTAB-7014:input_flagSMARCAD1
broker name
ArrayExpress
cell line
PGK12.1
cell type
embryonic stem cell
common name
house mouse
disease
normal
genotype
SMARCAD1-3xFLAG
organism part
inner cell mass
sample name
E-MTAB-7014:input_flagSMARCAD1
strain
PGK x 129P2/Ola/Hsd

Sequenced DNA Library

library_name
input_flagSMARCAD1_s
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
Approximately 5x10e7 PGK12.1XX cells per 15 cm plate were cross-linked with 2 mM DSG (disuccinimidyl glutarate, Thermo Scientific) in PBS for 45 min at room temperature, washed three times with PBS, and formaldehyde (Polysciences) was added to a final concentration of 1 % for 10 min at room temperature. Formaldehyde was quenched using glycine (0.125 M). Mouse PGK12.1 XX ES cells were cultivated feeder-cell free on gelatin-coated dishes in Dulbecco's Modified Eagle Medium (DMEM, Life Technologies) supplemented with 15% fetal calf serum, 2 mM L-glutamine, 1x non-essential amino acids, 50 μM 2-mercaptoethanol, and 1000U/ml ESGRO leukemia inhibitory factor. Generation of FLAG-SMARCAD1 expressing PGK12.1 XX ES cells: The coding sequence of SMARCAD1 was amplified from mouse ES cell cDNA and inserted into a chicken beta actin promoter (CAG)-driven expression vector, with an N-terminal triple FLAG tag. Tagged wild-type Smarcad1 construct along with the empty FLAG vector control were transfected into PGK12.1 ES cells using Lipofectamine 2000 (Life Technologies), followed by 1.7 µg/ml puromycin selection and clonal expansion. Chromatin was eluted in 1% SDS, 100mM NaHCO3 and DNA purified by Qiaquick PCR cleanup columns (Qiagen). Library preparation was performed using the MicroPlex Library Preparation Kit v2 (Diagenode) according to manufacturer's instructions. Briefly: 7ng DNA was used for each sample. Adapter-ligated DNA was subject to 9 cycles of PCR amplification before size selection and DNA purification by AMPure XP beads (Agencourt).

Sequencing Platform

instrument_model
Illumina HiSeq 1500

mm10

Number of total reads
49561987
Reads aligned (%)
98.0
Duplicates removed (%)
8.3
Number of peaks
493 (qval < 1E-05)

mm9

Number of total reads
49561987
Reads aligned (%)
97.9
Duplicates removed (%)
8.3
Number of peaks
504 (qval < 1E-05)

Base call quality data from DBCLS SRA