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文献和实验PRODUCTION OF COMPLETELY ES CELL-DERIVED FETUSES BY AGGREGATION WITH TETRAPLOID
The technique described here is a slight modification (March 1997) of methods presented in: Nagy A., J. Rossant. 1993. Production of completely ES cell-derived fetuses. In : Gene Targeting: A Practical Approach. (ed. A. Joyner). IRL Press at Oxford
for four hours in the presence of colcemid (3.125 µl of 10 µg/ml colcemid (Gibco # 15212-012) per ml of medium). 2ml per 35 mm and 4ml per 60 mm. 3) Remove media and place it in a 15 ml Falcon tube. Wash with trypsin, and trypsinize with 1.5 ml trypsin.
ES and TS cell freezing/thawing
Needed: ES cell freezing medium (2x) 2x ES cell freezing medium should be made up fresh each time it is to be used, and should comprise freshly prepared 60% DMEM+, 20% FCS, and 20% DMSO (Sigma, Cat. No.D-5879). Freezing
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