Prep of DNA for electroporation

Frankster f.lee at utoronto.ca
Sun Nov 24 00:56:05 EST 1996



>BARRETT at RASCAL.MED.HARVARD.EDU wrote:
>: Dear fellow netters,

>: I have recently run across a problem that hopefully someone out there has also
>: run into.  Prior to electroporation, I precipitated a ligation mixture with
>: EtOH.  But then when I went to electroporate the DNA, I got arcing in the
>: cuvette-- apparently some of the agarose still present in the ligation mixture
>: came down with the DNA during precipitation.  My question is, how do you get
>: rid of this agarose? In my hands, phenol/chloroform extraction will only 
>: partially remove the agarose, is there something else I should try?

>: Thanks much,

>: Peter Barrett
>: barrett at a1.tch.harvard.edu

We always use CsCl2 purified maxi preps.  Other large scale
purifications (Qiagen, Monster, etc.) will work as well.  In all
instances making sure that most of maxi prep'd DNA is supercoiled.





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