Reuse of Quiagen columns
Hernan Espinoza
espinoza at cgl.ucsf.edu
Wed Sep 20 17:07:10 EST 1995
In Response to the post by jnbradsh at STUD.MED.CORNELL.EDU (John Bradsher)
about regeneration/reuse of Quiagen columns...
(for some reason my reader won't let me quote today, sorry)
I reuse Quiagen columns routinely. Basically, I run my clarified lysate
through once -> wash with QC -> elute with QF -> wash with a bit more QF
-> wash with QC -> more lysate -> repeat etc. I have tried this with up
to FOUR cycles without any obvious erosion of purity. I did find that
after three cycles I already HAD most of the plasmid DNA, so I normally
only use TWO cycles,these days. What does this prove? Not much. All I
did was follow supercoiled DNA on a EtBr stained gel and take some A260/
A280 readings, so I can't claim real quantitation.(vis a vis the state of
the column after the prep) PLUS all of this was done with one lysate
and afterwards the column got da boot.
That said, I agree that there must be a way to regenerate these columns,
so that they can be used multiple times (ie different plasmids ,etc.)
Does anyone out there know what the resin is, that should give us a
good start. -Hernan
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