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文献和实验. Avoid repeated freeze-thaw cycles. All enzymes, upon reconstitution, can be sterile filtered through a 0.22 µm pore size membrane. Generally most of the enzymes used in cell isolation procedures (except trypsin) can be directly
PRODUCTION OF COMPLETELY ES CELL-DERIVED FETUSES BY AGGREGATION WITH TETRAPLOID
of aggregation plate Materials: Dissecting microscope; Sterile tissue culture dishes (Easy Grip 35 x 10 mm, Falcon 3001-3); 1 ml syringe with 26 G 1/2 needle of KSOM medium; light mineral oil (EMBRYO TESTED) (e.g. Sigma: M8410);
(for commonly used E:T of 20:1), aliquot 50 µl into each well (triplicates), After 1 hour incubation, spin target cells at 1000 rpm for 5 min. Decant supernatant, resuspend in 40 ml R10 media. Spin again and resuspend in 50 x the original cell
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