IUBio

DNA Quality and Transfections - does it really make a difference?

George Rutherford gruther at bilbo.bio.purdue.edu
Mon Mar 1 15:47:14 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 work in a fly lab, that in addition to the usual P-element
transformation stuff also uses S2 cells (an immortal fliy line of some
kind) to express proteins. No one that I know of has ever used anything
but double-banded DNA for transfections. The reason for this may be the
requirement of using similar DNA for embryo injections; it noetheless
works well (Ca Phosphate method).

George



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