Lidstrom:Ultracentrifuge: Difference between revisions
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** [[image:tubes collapsed in ultracentrifuge.jpg|thumb|center|Tubes with only 1/2 mL liquid collapse in ultracentrifuge.]] | ** [[image:tubes collapsed in ultracentrifuge.jpg|thumb|center|Tubes with only 1/2 mL liquid collapse in ultracentrifuge.]] | ||
** It is hard to say how much is enough, but it seems to be about 1 mL. 1/2 mL '''definitely''' collapsed on [[User:Janet B. Matsen|Janet]] in 5/2013. | ** It is hard to say how much is enough, but it seems to be about 1 mL. 1/2 mL '''definitely''' collapsed on [[User:Janet B. Matsen|Janet]] in 5/2013. | ||
** We should be able to find smaller-volume tubes for our rotor. ([https://www.beckmancoulter.com/wsrportal/wsr/research-and-discovery/products-and-services/centrifugation/optima-max-xp/index.htm link]) | |||
** You probably won't be able to get any sample out of a collapsed tube. | ** You probably won't be able to get any sample out of a collapsed tube. | ||
** When they collapse it is very tricky to get them out. Needle nose pliers are probably your best bet but it can still be very hard. | ** When they collapse it is very tricky to get them out. Needle nose pliers are probably your best bet but it can still be very hard. | ||
Revision as of 17:07, 26 May 2013
Back to Protocols
Our System
- Ultracentrifuge = Beckman Optima MAX Ultracentrifuge
- Rotor = MLS-50 from Beckman Coulter
Basics
- If you desire 4oC during your spin cycle, you must allow the machine to cool to that point before it will allow you to press start.
- If you use too small of a volume in the ultracentrifuge tubes, they will collapse!
Tubes with only 1/2 mL liquid collapse in ultracentrifuge. - It is hard to say how much is enough, but it seems to be about 1 mL. 1/2 mL definitely collapsed on Janet in 5/2013.
- We should be able to find smaller-volume tubes for our rotor. (link)
- You probably won't be able to get any sample out of a collapsed tube.
- When they collapse it is very tricky to get them out. Needle nose pliers are probably your best bet but it can still be very hard.
- If the ultracentrifuge won't start and won't even try to start, open the door, lift the rotor up, place it back down, let it cool back down to your desired temp, and hold start.
Example Protocols
- To spin down the membranes and membrane-bound proteins in a crude extract, centrifuge for 45,000 RPM at 4oC for 1 hour. With our MLS-50 mushroom-shaped rotor, this is 163,000xg at raverage (108,000xg at rminimum).
