2 replies to this topic

[Myco Rax]

Hello all,
I wanted to transfect the HEk cells with lentiviral vector.
Before addition of the transfection complex I forgot to treat the cells with chloroquine.
what is the purpose of chloroquine in transfection? Can anyone please tell me whether the transfection has taken place, as i forgot to add chloroquine?


Thanks in advance.

Edited by Myco Rax, 13 October 2011 - 11:14 PM.

[leelee]

It seems that chloroquine greatly improves transfection efficiency (some report 2 fold increase, others more). I couldn't find how it works, it appears to not be known, but I didn't look very thoroughly and the only paper I found about it was from 1996. This only took me five minutes to find out with google so I'm sure you'll be able to find out more if you have a good look.

So yes, you may have had successful transfection.

[dpo]

Chloroquine is an inhibitor of the lysosomal degradation of the DNA which is taken up by the cells, so as leelee said, transfection should have been successful, albeit at a slightly lower level than if you would have added the chloroquine. In my normal transfections, I never add chloroquine and get high efficiencies nevertheless.  Only when I'm making virus, I add it (although I must admit I never tested virus production without chloroquine).