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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
Cell counting with an hemacytometer.
Inc. Garden grove CA 92841 Hemacytometer Fisher scientific No 02671 5 Dilute your cell sample in Trypan Blue dye exclusion medium. Dead cells will be blue, live cells will be birefringent. Suggested dilution 1:20 (5µl cell suspension in 95 µl Trypan Blue
with pasteur pipet. Resuspend cells in 0.5 to 0.75 ml of fresh fix and gently draw suspension up in a pasteur pipet(will make 4 to 5 slides). 9) Dip slides (plain, pre-cleaned slides (Fisher #12-549) labelled with etching tool) in water, leaving a puddle
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