what speed to centrifuge down agarose bead? - (Nov/28/2006 )
I saw some protocals said at low speed to avoid crashing the bead, some just at full speed, some pulse at full speed (ie 5 sec at full speed). I did at low speed ,feel the bead is not packed enough, so it's hard for me to aspirate the sup.
I spin the amersham one at 12000g 30s. It's OK
I did the same with beads from an other provider, but it was bad for them. they were compacting.
use the protocol given by the provider.
I agree with missele - if there is no protocol of the manufacturer be defensive and try to restrict rpm to the lowest but efficient speed;
beside prevention of the breaking of beads, I do not know if a rapid pelleting may have effects on the specific binding, I mean if a high g-number would force to part some of the binding partners...
any suggestions to this point??
using M2 agarose beads from sigma, 2000rpm in a swing out rotor for 30'' do the job.
centrifuge at very low speed (2000) and if its impossible to remove super increase very gradually until you are able to. (pray not to crush the beads!)