reusing plasmid maxiprep columns

theodorn at medlib.georgetown.edu theodorn at medlib.georgetown.edu
Thu Apr 9 09:45:41 EST 1998


In article <KIvqTHA634K1Ewcp at demon.co.uk>,
  "Dr. Duncan Clark" <duncan at genesys.demon.co.uk> wrote:
>
> In article <6ge8if$fkh at mserv1.dl.ac.uk>, Wolfgang Schechinger
> <wgschech at med.uni-tuebingen.de> writes
> >I noticed that plasmid maxi columns e.g. nucleobond
> >(clontech/macherey-nagel) or those from quiagen may be re- used
> >several times.
> >currently, I use them only for the same plasmid (from XL-1 blue) and
> >wash them with plenty of water after eluting the plasmid. has anyone
> >tested how long the columns will leak plasmid, e.g. by repeating the
> >prep without bacterial lysates and checking for plasmids in the elute
> >by transforming aliquots ef the elute into bacteria? In other words,
> >does anyone dare to use the columns for different preps? what
> >lifetime of the columns may be expected?
> >I (and probably many others out here) would appreciated any data or
> >even references on that!
>
> Given that they are anion exchange resins ie QAE or similar then it
> should be possible to wash off all the junk that is still bound after
> the plasmid has been eluted. Water will not do. Try much higher salt or
> maybe even 7M Urea and 1%SDS as that is a real good cleaner on dye
> columns. NaOH may sterilise the columns sufficiently for other plasmid
> preps.
>
> Duncan

Although, if the matrix for the column is silica-based, then NaOH will
dissolve the resin.  What is the pH of the elution buffer?  If it is very
high, then that might limit the number of times the column could be used, if
it is a silica resin.

I'm getting some deja-vu; hasn't this topic been discussed here a couple of
years ago?

Also, for anion-exchange resins in general, you would want to avoid SDS since
it will bind to the column.

Nick Theodorakis
Georgetown University Medical Center

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