Ulrich Maier maieru at uni-muenster.de
Fri Aug 29 02:52:21 EST 1997

CANTOLUPO at GENVEC.COM (Lou Cantolupo) wrote:

>I recently column purified a fairly large plasmid (about 40kb) from a
>column.  After elution, pption/wash, and drying, I now cannot get it
>back into solution.  What I've tried:

>1.  Expected yield is about 20ug.  I dried for about 5 min at RT (until
>pellet was slightly translucent) and added 40ul of TLE (10mM Tris 8.0,
>0.1mM EDTA).  Let set at RT for 1 hour, did not dissolve.

>2.  Transfered to 65oC water bath for one hour...no dissolution.

>3.  Increased volume to 100ul with more TLE, set at 65oC for one hour. 
>Still there.

>4.  Incubated at 37oC o/n.  Has not gone into solution.

I had the same problems with genomic DNA. Try increasing the volume of
TE to 200 or 500µl, dissolve the plasmid over night at room
temperature or 4°C with gentle agitation of the tube.

You can reduce the volume with a speed vac after cutting the DNA.

Hope this helps,

Uli Maier

More information about the Methods mailing list