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Protocol for transfection of mES using lentivirus - (Feb/07/2008 )

Does anybody has the experience in transfection of mES cells (fedder dependent) using lentivirus?


I think you mean transduce.

You make the lentivirus in a cell line, such as 293T. You then take the supernatant, concentrate (this is optional), and then transduce the cells of choice. These are then sorted for GFP expression (assuming you have GFP).
There are quite a few papers out there on this subject. Also try a search in this forum.



Do you have a protocol for the transducion?


For primary neurons, we add the lentivirus and leave it for a few hrs and then change media. Check literature for mES .