Transfection with DEAE Dextran - (Oct/12/2007 )
I want to transfect CHO cells with DEAE Dextran. I found a manual by google, but I am not sure about it. Normaly you sead cells the day before transfection, but in this manual they sead them after transfection. So my question; what do you do?
Ah, DEAE-dextran, for some reason I hate it. There are 2 ways to perform the transfection on adherent cells:
The first protocol involves applying the dextran/DNA mix directly to the cells on the dish for a short period of time, usually in the presence of chloroquine, followed by a DMSO or glycerol "shock". This is the techinique I am familiar with.
The second method involves exposing trypsinized cells in solution to the DNA/dextran mix. I have never tried this, but have read it's for difficult to transfect cells that give poor results with the first method.
I have two different protocols for transfecting cells on the plate and one for cells in suspension. One adherent cell protocol my PI gave me and used throughout her post-doc-and is the one I use and the other protocol comes from Short Protocols in Molecular Biology. Let me know if you would like one or all of these protocols.