SAMN35750883
External Id
Developmental Genetics, Max Planck Institute for Molecular Genetics
INSDC center name
2023-10-31T00:00:00Z
INSDC first public
2023-10-31T15:29:08.500Z
INSDC last update
SRS17981195
INSDC secondary accession
live
INSDC status
Generic
NCBI submission model
Generic.1.0
NCBI submission package
SRS17981195
SRA accession
genetically modified F1G4 (C57BL/6N x SV129) mES cells
cell line
in vitro generated notochord progenitor-like cells
cell type
missing
collection_date
TRE::Foxa2-IRES-EGFP (Rosa26 locus); T::H2B-mCherry-T2A-irTA (BAC); Noto::H2B-mTurquoise (BAC)
genotype
missing
geo_loc_name
Mus musculus
http://purl.obolibrary.org/obo/NCBITaxon_10090
organism
FACS sorted Noto+/T+ cells from in vitro differentiated cultures
source_name
FACS sorted Noto+/T+ cells from in vitro differentiated cultures
tissue
ChIP H3K4me1 NotoPdiff
title
Cells were washed 2x PBS and dissociated by trypsinization at 37°C for 10min. Trypsin/EDTA was quenched using a double volume of 5%BSA/PBS, resuspended and kept on ice until further procedure. All samples were immediately filtered (35µm mesh) and sorted on a FACS Aria II (Becton Dickinson) flow cytometer. Cells were sorted into PBS 5% BSA in BSA coated glass tubes. After sorting, cells were pelleted by centrifugation, transferred to 1.5 ml low binding tubes and washed in PBS containing 0.05% Triton X-100 to break the surface tension. Crosslinking was performed by adding 1/10th volume of crosslinking solution (11% formaldehyde, 100mM NaCl, 1mM EDTA pH8, 0.5mM EGTA pH8, 50mM Hepes pH7.8) to cells resuspended in 500µl PBS for 10 minutes at room temperature with agitation. The crosslinking reaction was quenched with the addition of 1/10th volume of 2.5M glycine and 5 minutes incubation. Pellets were washed twice using cold PBS, containing 0.05% Triton X-100 to break the surface tension. Cells were pooled in aliquots of ~0.6x106 in Eppendorf tubes, snap frozen and stored at -80°C until sonication.
treatment