Re: Reconstitution of CNBr fragments
Ulf Hellman (ulf.hellman@licr.uu.se)
Tue, 12 May 1998 16:36:43 +0300
Hi Dougie,
Reconstitution of CNBr fragments? Depends on what you want to do with them.
If you want to isolate by e. g. gelfiltration, you should not dry them at
all, but keep them in the fragmentation solvent (70 % formic acid?); then
run the fragment mixture in a column equililbrated in the same solvent
(Superdex 75 should be ideal here).
If your fragments are dry, and you want to get N-terminal sequence of them,
I would (easily) dissolve them in "sample buffer" and run SDS-PAGE in a
tight gel. Then after blotting and Coomassie staining, cut out and stick
the PVDF pieces into the sequencer.
For other purposes, I also await info on the "Super Solvent"!
/Ulf
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Ulf Hellman, Ph. D.
Ludwig Institute for Cancer Research :
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