Have fresh ES media and a 24-well plate with enough wells for your targeted clones with feeders prepared.
1. Take the 96-well plates from the -80 C freezer and place directly into the 37oC incubator. Allow the cells to thaw completely. (~10-15 min.)
2. After the plate has thawed, remove the entire contents of the 96-well plate and place in one well of a 24-well plate.
3. Pipette up and down 2-3X to dissociate and spread the cells out.
4. Do not feed cells until you begin to see colonies growing in you wells. This could take 2-3 days.
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