probe separation

Thibaud Le Mouel lemouel at antibes.inra.fr
Thu Jul 18 01:34:33 EST 1996


>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

I use Sephadex G-50 and 2 ml syringe : prepare G-50 in TE buffer, take a
syringe and put in some glass wool and then 2 ml of G-50, put your syringe
in a 13 ml tube with a 1,5 ml eppendorf (without its cap) in the tube turn
at 1600 rpm during 4 minutes, wash your column with TE and turn at 1600 rpm
during 4 minutes (repeat 2 times). Then you put your probe on the column
and turn it 4 minutes at 1600 rpm. You have your clean probe in the
eppendorf and the unincorporated nucleotides are still in the column.

Good luck

Thib

=========================================================================
******** **       *       * *********************************************
******** **       **     ** * Th. Le Mouel  <lemouel at antibes.inra.fr.>  *
   **    **       ***   *** * I.N.R.A.                                  *
   **    **       **** **** * 123, bld F. Meilland   B.P. 2078          *
   **    ******** ** *** ** * 06606 ANTIBES CEDEX    FRANCE             *
   **    ******** **  *  ** *********************************************
=========================================================================
"Up the Irons..... Down to the pub !!"





More information about the Methods mailing list