ultracentrifuge problems

Georg Kroeger a03m at biologie.uni-bremen.de
Thu Dec 12 14:25:59 EST 1996


L.Force at botany.uq.edu.au (Lesleigh Force) wrote:

>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 placed the tubes in the 
>rotor.

>Any ideas on how to prevent the tubes collapsing?

>Thanks.

>Lesleigh Force
>PhD student
>L.Force at botany.uq.edu.au


Hi Lesleigh,

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!

Cheers

Georg




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