Hi everyone,
I'm having problems with ultracentrifuge tubes collapsing/breaking at
the top during a run. I'm using a Sorvall ultracentrifuge OTD75B with
T-875 fixed angle rotor and 11.5ml ultracrimp tubes (cat. no. 03987).
I'm trying to separate protein/pigment complexes of isolated thylakoids
using a 9ml 0.125-1.0M sucrose gradient made up in 40mM MES-AMPD pH6.5
(MES = 2[N-morpholino]ethanesulfonic acid, AMPD =
2-amino-2-methyl-1,3-propanediol), 1% B-dodecyl-maltoside and 0.1mM
phenylmethylsulfonylfluoride. The tubes were spun at 4 c at 49000rpm
for 20 hours. No air layer was visible when I place them in the rotor.
Any ideas on how to prevent the tubes from collapsing?
Thanks.
Lesleigh Force
PhD student
L.Force at botany.uq.edu.au