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文献和实验ES Cell Culture and Manipulation
CAN be stored short term (a few moths) at -20°C. Titering LIF Plate 1 x 104 feeder-free ES cells in each of the wells of a gelatinized 6-well in ES media without LIF. At plating add LIF at 1:100, 1:500, 1:1000, 1:2500, 1:5000 and 1:10,000
ES Cell Culture and Manipulation
and harvest a second batch of LIF the next day. Throw these cells out when you are done as, they have now been confluent for ~2 days. LIF conditioned media CAN be stored short term (a few moths) at -20°C. Titering LIF Plate 1 x 104 feeder
media and wash once with PBS. 2) Freeze down the 3 x gelatinized only plate at –20 °C for DNA prep. 3) For the 2 x plates with feeder cells, add 25 µl trypsin STV and incubate at 37 °C for 5 minutes. 4) Disaggregate cells with 100 µl
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