DNA WON'T DISSOLVE!!
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.
>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,
More information about the Methods