G4 ROSALUC B12 cell line

Source: Mouse

Type: Embryonic stem (ES) cells

Cell line designation: G4 ROSALUC B12

Other designation: G4 ROSALUC ES

Other culture collection numbers: /

Containment level: This cell line has beenassigned the containment level ‘Class 1’

following the European Directive 2009/41/ECon the contained use

of genetically modified organisms, and itsupdates

Medium:

? 500 ml bottle Knockout DMEM

? 90 ml serum

? 6 ml penicillin/streptomycin 100X

? 6 ml beta-mercaptoethanol 100X

? 6 ml non-essential amino acids 100X

? 6 ml glutamine 100X

? recombinant mouse LIF 2000 units/ml

For selective medium, use puromycin at 1μg/ml

Selection marker: puromycin

Cultivation temperature: 37°C

Original reference: Haenebalcke et al.,Stem Cell Rev. 6 (2013), 774-785 [PMID:

23877658]

Related reference: /

Genotype: Heterozygous

Phenotype: /

Quality controls:

? Mycoplasma and other contaminations:negative

? Karyotyping by chromosome counting: 78%euploid

? (C57Bl6/Nx129S6/SvEvTac)F1 confirmationby PCR with

microsattelite D1 mit 303 (B6 = 142 bp;129S6 = 118 bp)

? PCR confirmation of presence of RMCEcassette: ES-cells

heterozygous for RMCE cassette

Further info: ‘B12’ in the name refers tothe clone number.

Culture recovery and preservationinstructions

Recovery: startup in a 6-well withinactivated feeders after washing.

Long-term preservation: cryopreservation(at least at -120oC)