David L. Haviland, Ph.D.
dhavilan at IMM2.IMM.UTH.TMC.EDU
Tue Aug 5 09:21:52 EST 1997
At 17:27 8/4/97 -0400, Molecular Pathology Lab wrote:
> I was wondering in anyone has a protocol for doing a G-50 spun column
>using a microcentrifuge for seperating unicorporated from labled probe.
>I have used the 1mL (tuberculin) syringe method, but I don't have access
>to a clinical centrifuge, but I do have access to an old
>microcentrifuge. Any help that you can give me will be greatly
>appreciated, and you can either send it to my e-mail or to the news
Yes, we do all the time. Here how... We use the Fischer Brand spin columns
#11-387-50 and we buy both G50 and G25 from Pharmacia. The sepharose is
made per direction as resuspended final 50:50 with TE. Our standard
columns are 3-4 mls. We spin in a IEC but a JT6 or equivalent should
sufice. A prep spin of 2 mins at 2000 rpm to remove excess water is done,
then the labeled sample is applied (tube swicth of course) then spun again
4 mins at 2000 rpm. Done. Many times, I;ve prepped G25 with DEPC water
and used the column RNAse free.
Hope this helps,
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