Purification of large amounts of restriction fragments

Lisa Kueltzo kueltzo at ukans.edu
Thu Aug 3 14:04:19 EST 2000


Greetings all,

I would like to purify a series of DNA restriction fragments in the
range of 100 to 1000 bp (mg or possibly 100s of ug each of several
sizes).  While the literature as well as catologs from TosoHaas and
Pharmacia provide several methods for analytical separations of such
fragments I have not had much luck in finding any information regarding
isolation of such large amounts.  Any replys regarding the feasibility
of such a high yield as well as suggestions regarding where to start
would be greatly appreciated.  Initial attempts will be with
chromatography resins used for the analytical columns (ion-exchange and
gel filtration).

Starting materials such as DNA and restriction enzymes are not the
problem for us.  Isolation of these fragments is the part causing all of
the head scratching.

Thanks for your comments and suggestions,

Chris


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