NextSeq 500 sequencing; Genome wide analysis of STAT3-mediated transcription and binding sites in mouse female germ stem cell (ChIP-seq)
Sample information curated by ChIP-Atlas
Antigen
Antigen Class
No description
Antigen
NA
Cell type
Cell type Class
Gonad
Cell type
Germline stem cells
NA
NA
Attributes by original data submitter
Sample
ENA first public
2017-12-20
ENA last update
2017-11-10
ENA-CHECKLIST
ERC000011
External Id
SAMEA104376754
INSDC center alias
School of Biomedical Engineering, Shanghai Jiao Tong University
INSDC center name
School of Biomedical Engineering, Shanghai Jiao Tong University
INSDC first public
2017-12-20T17:02:49Z
INSDC last update
2017-11-10T15:10:06Z
INSDC status
public
Submitter Id
E-MTAB-6163:Sample 2
age
5
broker name
ArrayExpress
cell line
Female germline stem cell line
common name
house mouse
disease
normal
genotype
wild type genotype
organism part
female gonad
sample name
E-MTAB-6163:Sample 2
sex
female
strain
C57BL/6
Sequenced DNA Library
library_name
Sample 2_s
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
FGSCs at passage number 35–45 were cultured on gelatinized plates with minimum essential medium (MEM)-α (Gibco) containing 10% fetal bovine serum (Gibco), 1mM non-essential amino acids (Gibco) , 1mM sodium pyruvate (Gibco), 2mM L-glutamine (Gibco), 0.1mM β-mercaptoethanol (Sigma-Aldrich), 10ng/ml mouse epidermal growth factor (Pepreotech), 20ng/ml mouse leukemia inhibitory factor (Pepreotech), 10ng/ml human glial cell line-derived neurotrophic factor (Pepreotech), 10ng/ml human basic fibroblast growth factor (Pepreotech), and 100units/ml Penicillin-Streptomycin Solution at 37 °C in a 5 % CO2 atmosphere as previously described (Kang Z, et al. Nat Cell Biol. 2009). cells were covalently cross-linked by incubating cells in 1 % formaldehyde (Sigma-Aldrich) for 10 minutes at room temperature and incubated with 125 mM glycine for 5 minutes to quench cross-linking reaction. The fragmented chromatin fragments were immunoprecipitated with Protein A+G Magnetic beads (Millipore) coupled with anti-STAT3 antibody (13-7000, Invitrogen). After reverse crosslinking, ChIP and input DNA library were constructed with a NEBNext Ultra II DNA Library Prep Kit for Illumina (E7645, NEB).