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文献和实验Freezing and Thawing of Mammalian Cell Lines
10e6 cells/0.5 mL freezing medium. Aliquot in cryo-vials 0.5 mL/vial. Place vials upright in a styrofoam box and cover well. Place in a -70 °C for 24 hours. Place vials in a wand and put in a liquid nitrogen container. Thawing
Freezing and Thawing of Mammalian Cell Lines
% FCS, 70% media that you used to grow the cells such as RPMI or DMEM) at a concentration of 2 X 10e6 cells/0.5 mL freezing medium. Aliquot in cryo-vials 0.5 mL/vial. Place vials upright in a styrofoam box and cover well. Place in a -70 °C for 24
. To make bacteria for seeding plates, use a flamed wire loop to pick a single OP50 colony into a 100 ml bottle of B broth. Set the bottle (no shaking) at 37° overnight. It should be cloudy the next day. This bottle can be stored at 4° and used for ~3 months
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