SAMN35633307
External Id
University of Cambridge
INSDC center name
2023-06-30T00:00:00Z
INSDC first public
2023-06-30T17:41:01.803Z
INSDC last update
SRS17897433
INSDC secondary accession
live
INSDC status
Model organism or animal
NCBI submission model
Model.organism.animal.1.0
NCBI submission package
SRS17897433
SRA accession
Embryonic day 13.5
age
MEF-D
cell_line
Primary mouse embryonic fibroblasts (MEFs)
cell_type
2022-06
collection_date
E13.5
dev_stage
Dnmt3aflox/flox3bflox/flox
genotype
United Kingdom: Cambridge
geo_loc_name
Primary MEFs were established from Dnmt3aflox/flox3bflox/flox E13.5 embryos and grown at 37°C and 5% CO2 in high glucose DMEM GlutaMAX™ (ThermoFisher, cat no. 31966021) supplemented with 10% FBS (Gibco, 11550356) and 1% penicillin/streptomycin solution and passaged with trypsin/EDTA. To induce the knockout of DNMT3A/3B, 1x106 cells were plated and the next day washed with PBS and media was replaced with 5ml of DMEM:PBS (1:1) with 50µl TAT-CRE recombinase (2-3µg/µl) and incubated at 37°C for 8 hours. Following the TAT-CRE recombinase treatment, cells were washed three times with PBS and the original growth media was replaced. The TAT-CRE recombinase treatment was carried out again 48 hours following the start of the initial treatment. Cells were collected for protein and DNA/RNA extraction 72 hours after the second TAT-CRE recombinase treatment.
growth_protocol
Mus musculus
http://purl.obolibrary.org/obo/NCBITaxon_10090
organism
cell culture
sample_type
female
sex
C57BL/6
strain
Day 13.5 embryos
tissue
Primary MEF cells, population D, DNMT3A/3B DKO (+ TAT-CRE)
title
TAT-CRE treated
treatment