START QUOTE:
If you like the results you get from CentraSep columns, you can always
"roll your own" by re-packing them with DNA-grade Sephadex G-50. The
easiest way to do this is to pour out the Sephadex from a prepacked
column into a microfuge tube. Mark the level of Sephadex in the tube
with a Sharpie and cut off the top of the microfuge tube to the line
with a sharp razor blade. You now have a "scoop" for measuring out the
correct quantity of Sephadex to re-pack (glue on a pipet tip as a
"handle" if you like, or just use a clamp or forceps as the handle). To
re-use the columns, simply let the Sephadex dry out in a used column and
dump out the dry pellet (it shrinks and falls right out when dry). Then
rinse the columns in water, sonicate for a few minutes in distilled
water, and allow them to dry. Re-pack with dry Sephadex using your
homemade scoop and you're all set for the next round. I'm sure the
folks at Princeton Seps won't like it, but it works fine and its much
cheaper. Cheers.
END QUOTE!
It works very well for our low level of sequencing and I calculate the
marginal price in materials at about 10 cents / column. Some people only
re-use the columns 2 or 3 times but we continue to get good results after
multiple re-uses. The only added labor is the sonication cleaning step
which we have not optimized so in our hands works well but is labor
intensive. For the Sephadex we use Pharmacia Fine DNA grade Sephadex G-50
listed in their automated sequencing section (cat NO 17-1573-01).
good luck
-- Chris Shaffer Box 1229; Biology Dept Washington University One Brookings Dr St. Louis MO 63130
---------- >From: Phyllis Spatrick <pspatric@sunspot.ummed.edu> >To: Recipients of ABRF List <abrf@aecom.yu.edu> >Subject: BigDye Cleanup >Date: Thu, Oct 21, 1999, 8:26 AM >
> Hello Everyone > We recently switched to using big dyes from the TaqFS. We are finding > that our sample cleanup method of G50 beads packed into a spin column is > not removing enough of the dye now. We tried a Princeton separations > column which was much better, but that seemed akward and it is expensive. > What do other labs use? Any Suggestions? > Phyllis > > > Phyllis Spatrick > University of Massachusetts - Worcester Campus > Phyllis.Spatrick@ummed.edu >