First, prepare DMEM serum medium
1. Take the FBS from the refrigerator at -20 °C one day in advance, and put it in a refrigerator at 4 °C;
2. FBS and other components to be added (such as sodium pyruvate, antibiotics, etc.) are added to the DMEM medium in proportion to the cells, and placed in a refrigerator at 4 °C.
The preparation ratio is as follows:

DMEM (High glucose) / DMEM medium (high sugar)

Glutamine

50ul

Non-essential amino acid

500ul

Sodium pyruvate

500ul

B-mercaptoethanol

Double antibody

50ul

serum

7.5ml

LIF

5ul

Second, ES cell culture medium
DMEM+15% FBS

2-mercaptoethanol

5.5uM

1000X

L-glutamine

2mM

100X

LIF

1000U/mL

10000X

Non-essential amino acid

100uM

100X

Double antibody

100X

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