probe separation

JLBlackwell jblac at vetmed1.vetmed.ufl.edu
Wed Jul 17 14:27:19 EST 1996


In article <199607171748.NAA25659 at photon.magnus.acs.ohio-state.edu>,
jprei001 at POSTBOX.ACS.OHIO-STATE.EDU (Johanna Preiszner) wrote:

> Hi,
> Do you have a good method for preparation of Sephadex G50 minicolumns? I
> want to use it to separate ramdom-prime labelled DNA and 32P-dCTP by
> centrifugation, avoiding the usage of long columns. But I could not find the
> proper material for bottom-plug, and the Sephadex flows out if I put it to
> the centrifuge.
> Any good idea is greatfully acknowledged.
> Hanna  

We use 1 ml syringes, and use the cotton plug from a serological pipet
(always available!). Just push the cotton in, tamp it down, put in your
Sephadex, place the syringe in a 14 ml polypropylene snap-cap tube (minus
the cap) and spin out the buffer to make the column. We put a screw-top
1.5 ml tube into a fresh 14 ml snap cap, place the syringe into it, load
the probe and spin. The probe is caught in the screw top, which can be
removed with forceps. All very cheap lo-tech, but very effective.

JB



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