help! how to pack a sepharose column

David J. Meyer ez005587 at
Fri Apr 30 20:13:09 EST 1993

nexus at (hUnix) writes:
: Don't let the column dry!
: Arseny Markov
: Virology/Oncology
: ex-KSU research fellow
: -- 

	I just thought I would point out the reason why you don't let the
column run dry, since this is usually not made clear (although always stated)!
If the level of buffer in the column runs below the top of the bed, air
will enter in its place. The air between the beads is not usually chased
down the column when more buffer is added; instead, the buffer usually
follows the easiest path down the column, and this does not necessarily
(or usually) involve the entire cross-sectional area of the column. This
effectively decreases the bed volume of your column. If the coulumn 'runs
dry,' it must be reequilibrated and repacked.

	Good luck!

David J. Meyer
djmeyer at

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