DEAE-dextran transfection/divalent cations

N.P.Dantuma at biol.ruu.nl N.P.Dantuma at biol.ruu.nl
Sun Jun 2 15:26:13 EST 1996


Hi,

COS cells can be transfected according to the protocol of Cullen (Meth. 
Enzym. 152: 684-704) by incubating associated cells with DNA-DEAE-
dextran complexes in PBS without Ca and Mg. This DEAE-dextran transfection 
works nicely and the efficiency is high.

However, I noticed that many cells dissociate during the 30 min incubation 
with the DNA transfection mix when the Ca and Mg are omitted (as expected 
since COS cells require divalent cations for attachment).

Am I right that Ca and Mg have to be omitted according to this protocol (it 
is not clearly indicated)?

What is the reason? Does it disturb transfection?

Is it worth trying to transfect in the presence of Ca and Mg?

Suggestions/answers preferable by e-mail. Thanks.

Nico Dantuma



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