deficient) overnight
at 37°C. Dilute the cells 1/50 in 1% glucose, 1.25 µg/mL thiamine, 0.5 µM
thymidine, 1 mM MgSO4 and 20 µM EdU in M9 minimal media. Grow this culture
at 37°C for 24 hours in the dark
deficient) overnight
at 37°C. Dilute the cells 1/50 in 1% glucose, 1.25 µg/mL thiamine, 0.5 µM
thymidine, 1 mM MgSO4 and 20 µM EdU in M9 minimal media. Grow this culture
at 37°C for 24 hours in the dark
deficient) overnight
at 37°C. Dilute the cells 1/50 in 1% glucose, 1.25 µg/mL thiamine, 0.5 µM
thymidine, 1 mM MgSO4 and 20 µM EdU in M9 minimal media. Grow this culture
at 37°C for 24 hours in the dark
deficient) overnight
at 37°C. Dilute the cells 1/50 in 1% glucose, 1.25 µg/mL thiamine, 0.5 µM
thymidine, 1 mM MgSO4 and 20 µM EdU in M9 minimal media. Grow this culture
at 37°C for 24 hours in the dark
deficient) overnight
at 37°C. Dilute the cells 1/50 in 1% glucose, 1.25 µg/mL thiamine, 0.5 µM
thymidine, 1 mM MgSO4 and 20 µM EdU in M9 minimal media. Grow this culture
at 37°C for 24 hours in the dark
deficient) overnight
at 37°C. Dilute the cells 1/50 in 1% glucose, 1.25 µg/mL thiamine, 0.5 µM
thymidine, 1 mM MgSO4 and 20 µM EdU in M9 minimal media. Grow this culture
at 37°C for 24 hours in the dark
deficient) overnight
at 37°C. Dilute the cells 1/50 in 1% glucose, 1.25 µg/mL thiamine, 0.5 µM
thymidine, 1 mM MgSO4 and 20 µM EdU in M9 minimal media. Grow this culture
at 37°C for 24 hours in the dark
deficient) overnight
at 37°C. Dilute the cells 1/50 in 1% glucose, 1.25 µg/mL thiamine, 0.5 µM
thymidine, 1 mM MgSO4 and 20 µM EdU in M9 minimal media. Grow this culture
at 37°C for 24 hours in the dark
deficient) overnight
at 37°C. Dilute the cells 1/50 in 1% glucose, 1.25 µg/mL thiamine, 0.5 µM
thymidine, 1 mM MgSO4 and 20 µM EdU in M9 minimal media. Grow this culture
at 37°C for 24 hours in the dark
deficient) overnight
at 37°C. Dilute the cells 1/50 in 1% glucose, 1.25 µg/mL thiamine, 0.5 µM
thymidine, 1 mM MgSO4 and 20 µM EdU in M9 minimal media. Grow this culture
at 37°C for 24 hours in the dark
deficient) overnight
at 37°C. Dilute the cells 1/50 in 1% glucose, 1.25 µg/mL thiamine, 0.5 µM
thymidine, 1 mM MgSO4 and 20 µM EdU in M9 minimal media. Grow this culture
at 37°C for 24 hours in the dark
deficient) overnight
at 37°C. Dilute the cells 1/50 in 1% glucose, 1.25 µg/mL thiamine, 0.5 µM
thymidine, 1 mM MgSO4 and 20 µM EdU in M9 minimal media. Grow this culture
at 37°C for 24 hours in the dark
deficient) overnight
at 37°C. Dilute the cells 1/50 in 1% glucose, 1.25 µg/mL thiamine, 0.5 µM
thymidine, 1 mM MgSO4 and 20 µM EdU in M9 minimal media. Grow this culture
at 37°C for 24 hours in the dark
deficient) overnight
at 37°C. Dilute the cells 1/50 in 1% glucose, 1.25 µg/mL thiamine, 0.5 µM
thymidine, 1 mM MgSO4 and 20 µM EdU in M9 minimal media. Grow this culture
at 37°C for 24 hours in the dark
deficient) overnight
at 37°C. Dilute the cells 1/50 in 1% glucose, 1.25 µg/mL thiamine, 0.5 µM
thymidine, 1 mM MgSO4 and 20 µM EdU in M9 minimal media. Grow this culture
at 37°C for 24 hours in the dark
deficient) overnight
at 37°C. Dilute the cells 1/50 in 1% glucose, 1.25 µg/mL thiamine, 0.5 µM
thymidine, 1 mM MgSO4 and 20 µM EdU in M9 minimal media. Grow this culture
at 37°C for 24 hours in the dark
deficient) overnight
at 37°C. Dilute the cells 1/50 in 1% glucose, 1.25 µg/mL thiamine, 0.5 µM
thymidine, 1 mM MgSO4 and 20 µM EdU in M9 minimal media. Grow this culture
at 37°C for 24 hours in the dark
deficient) overnight
at 37°C. Dilute the cells 1/50 in 1% glucose, 1.25 µg/mL thiamine, 0.5 µM
thymidine, 1 mM MgSO4 and 20 µM EdU in M9 minimal media. Grow this culture
at 37°C for 24 hours in the dark
deficient) overnight
at 37°C. Dilute the cells 1/50 in 1% glucose, 1.25 µg/mL thiamine, 0.5 µM
thymidine, 1 mM MgSO4 and 20 µM EdU in M9 minimal media. Grow this culture
at 37°C for 24 hours in the dark
deficient) overnight
at 37°C. Dilute the cells 1/50 in 1% glucose, 1.25 µg/mL thiamine, 0.5 µM
thymidine, 1 mM MgSO4 and 20 µM EdU in M9 minimal media. Grow this culture
at 37°C for 24 hours in the dark
flask. After incubating 24 hours at 37 ºC, 5% CO2 and
95% relative humidity, the MEF medium was replaced with
fresh mESC medium (Merck Millipore, Cat. No. ES-101-B).
Flasks were seeded with mESCs (Merck
Sigma Prod. No. N-8129 or dissolve the contents of 1 mg vial of
-Nicotinamide Adenine Dinucleotide, Reduced Form, Disodium Salt, Sigma Stock
No. 340-101, in the appropriate volume of Reagent A. PREPARE