fragment isolation problem
svetlov at oncology.wisc.edu
Mon Nov 23 17:22:06 EST 1998
In article <737obm$pfn$1 at mserv2.dl.ac.uk>, "Francisco Villamon"
<fvillamon at hotmail.com> wrote:
: Hi all!
: I want to isolate one fragment to cloning.....the problem is the
: separation of one fragment about 2.4 kb from another one about 2.9 kb
: in an agarose gel. The difference beetwen both after running is so short
: making difficult the isolation by gel cutting.
: What can I do for separate more the fragments?
Run 0.6-0.8% gel until bromphenol blue runs off it.
If you still feel that they are too close for comfort, instead of cutting
the pieces of gel out insert DE-81 paper in front and behind the band of
interest and continue running untill the band is all transferred to the
front piece of DE-81 (the other one prevents contamination with larger
fragments). DNA then can be eluted from DE-81 in high salt buffer.
V. Svetlov, Ph. D.
McArdle Laboratory for Cancer Research
University of WI, Madison
1400 University Ave.
Madison, WI 53706
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