Maxiprep for transfection

simonmccallum at hotmail.com simonmccallum at hotmail.com
Sun Apr 25 15:09:12 EST 1999


In article <2CCB6CA5A8A5D211AFD700A0240AFD6C01052DD4 at MCDC-US-NEWS>,
  zje8 at cdc.gov (John R. McQuiston) wrote:
> Qiagen's Maxiprep (No Aff.)  called Qiatip 500 I think, is for producing
> DNA endotoxin free.  I would contact them.
>
> John
>
> In article <7f80dn$a84$1 at nnrp1.dejanews.com>, bli1997 at yahoo.com says...
> >
> >Hi, I tried to maxiprep a plasmid using the method in "Current protocol", but
> >it seemed part of the plasmid wes always resistant to digestion. Anybody
> >knows what was happenning there? Was it because of alkyline treatment? How
> >should I modify the method?
> >
> >Also, I was considering Wisard Maxiprep. Has anyone tried it for
> >transfection? I was told that it worked OK if you reextract it with phenol,
> >which removes the endotoxin that the Wisard resin leave. Anyone tried?
> >
> >Your advice is highly appreciated.
> >
> >-----------== Posted via Deja News, The Discussion Network ==----------
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>
>
john

Considering that correctly prepared miniprep DNA will cut to completion just
fine,  you're problem is almost certainly your maxiprep protocol, alkaline
lysis should  be finished when the resuspended solution becomes clear, this
usually happens  within a few seconds, maybe a minute if you've a lot of
bugs, I've had this digestion problem in the past, and reducing NaOH/SDS
incubation time did  the trick, also, rolling the centrifuge tube along the
bench for a few seconds is the maximum agitation required.

I've had no problems with the following kits: bigger prep, qiagen and wizard

simon

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