probe separation

[Thibaud Le Mouel]

>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

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