probe separation

David Winterbourne sghk100 at sghms.ac.uk
Thu Jul 18 08:33:38 EST 1996


In article <199607171748.NAA25659 at photon.magnus.acs.ohio-state.edu>, 
jprei001 at POSTBOX.ACS.OHIO-STATE.EDU says...

>Do you have a good method for preparation of Sephadex G50 minicolumns? ...
>But I could not find the proper material for bottom-plug ...

Cut out small discs from GF/C glass fiber filters. The discs should just fit a 
1ml syringe. You can make a re-usable punch to do this by cutting the tip off 
a 1ml Gilson at the appropriate distance.  Using the syringe plunger push two 
discs to the bottom of the syringe. Fill the syringe with Sephadex G50 in 
6xSSC (I just use gravity - this results in some dilution, but gives good 
recovery, alternatively centrifuge before applying sample).

A 50ml Falcon 2070 BlueMax can be made into a column holder: Cut a hole, just 
large enough to take the syringe, in the blue screw cap lid with a cork borer. 
Place a microfuge tube in the 50ml tube, put on the screw cap lid, insert the 
column so that the tip enters the microfuge tube, apply sample and spin 1min 
at 1000rpm.

The metal centrifuge bucket serves as a good screen to protect you from 
radiation when transferring between work station and centrifuge.

-- 
Dr. David Winterbourne
Department of Surgery
St George's Hospital Medical School, London SW17 0RE, England
Tel: 0181-725-5581   Fax: 0181-725-3594




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