DNA extractin from gel HELP

Panhead McNipper wblank at gpu.srv.ualberta.ca
Tue May 2 13:25:13 EST 1995


Beav -

I was having a similar problem with my pulsed-fields -- thought I had no
DNA, spent weeks trying to see a band on a gel.  Solution:  Don't look for a
band.
The large pieces of DNA will be broken into a smear of small ones which you
will be extremely lucky to see.  Instead, do a random prime label of your
final eluates (I assume that the Quiaex kit is similar to GeneClean), TCA
ppt them, and run through a scintillation counter.  I was surprised to find
I had enough DNA to use as a decent probe.

Good luck,
Wally

In Article <beavis-2704951603430001 at mac64_58.med.utah.edu>,
beavis at butthead.utah.edu (beavis) wrote:
>I've been trying for awhile now to extract a 30Kb piece of DNA from a
>pulse field gel with no success.  Initally I used B-agarase method
>following the protocol that came with the enzyme.  This did not work so I
>switched to Qiagens Quiaex11 kit and still with no success.  It is a 1%
>low melting point agarose gel and we use TBE as a buffer.  The gel used to
>test for presence of DNA  in the extract never shows a band.  The piece of
>DNA is prsent on the original pulse field gel at very low conc. but I use
>cybergreen instead of ethidium to stain as this is much more sensitive but
>i see nothing.  All help gratefully appreciated.

It's all much appreciated.  Out!



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