DNA Quality and Transfections - does it really make a differe nce?

Evans, Krista KEvans at lifetech.com
Wed Mar 3 16:26:50 EST 1999


In article <w.ingram-2502990923380001 at b-wainwright.cmcb.uq.edu.au>,
w.ingram at cmcb.uq.edu.au (Wendy Ingram) wrote:

> I recently had a discussion with a colleague who said that in their lab
> all the plasmid maxipreps used for mammalian cell transfections were
> further purified with cesium chloride prior to use. In our lab we have
> just been using plasmid straight out of Geomed Jetstar maxi kits (or
> occassionally nucleobond kits) for our transfections.
> 
> What I am keen to hear opinons on is whether additional purification steps
> are actually worth the added effort? Do transfection efficiencies increase
> significantly over DNA straight out of commercial maxi kits??? If
> purification really helps then what is the best way to do it?
> 

I have found that DNA quality may or may not have an effect on your
transfection efficiencies.  Some cell lines are more sensitive than others
to endotoxin levels.  Your transfection reagent may also be sensitive to DNA
preparation.  I've seen this first hand with triplicate samples.

Krista Evans




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