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文献和实验PRODUCTION OF COMPLETELY ES CELL-DERIVED FETUSES BY AGGREGATION WITH TETRAPLOID
tip) alternatively: Finger controlled pipette (Manostat tubing, yellow tip, scalpel blade); 9'' Pasteur pipettes; 70% Ethanol; Alcohol burner or Bunsen burner; Sterile plastic Petri dishes (100 x 15 mm); Sterile
RNA purification using TRizol--TRIZOL法提取RNA
Reagents and EquipmentsTRIzol Reagent (Life Technologies cat# 15596-026) or TRI reagent (Sigma cat # T-9424)DEPC (RNase free) water or 0.5% SDS solution in DEPC treated waterChloroform (Fisher )Isopropyl alcohol (2-Propanol) (Fisher)75% Ethanol
ES / MEF cell culture and electroporation of targeting construct
(40ug of clean linear DNA in 1 X TE @ 1 ug/ul for each electroporation). Mix the cells and DNA with a 200-1000 ul barrier tip trying not to create bubbles. Add 400 ul of the cell/DNA mixture to a BTX cuvette. Set Electroporator as follows: 500
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