a03m at biologie.uni-bremen.de
Thu Dec 12 14:25:59 EST 1996
L.Force at botany.uq.edu.au (Lesleigh Force) wrote:
>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 placed the tubes in the
>Any ideas on how to prevent the tubes collapsing?
>L.Force at botany.uq.edu.au
we had a similar problem doing CsCl purifications of DNA (lots of
ethidium etc. in the centrifuge). Check your crimper-device. The
strengh of closing the tubes has to be regulated regularly. Ours has a
metal chip (took us some time to find but was worth it!) to check the
tightness of the crimped shutter.
Hope this helps!
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