Lidstrom:Ultracentrifuge: Difference between revisions

From OpenWetWare
Jump to navigationJump to search
No edit summary
Line 7: Line 7:
== Basics ==  
== Basics ==  
* If you desire 4<sup>o</sup>C during your spin cycle, you must allow the machine to cool to that point before it will allow you to press start.
* If you desire 4<sup>o</sup>C 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!'''
* '''If you use too small of a volume (< half full) in the ultracentrifuge tubes, they will collapse!'''
** [[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.  
Line 17: Line 17:
== Example Protocols ==
== Example Protocols ==
* To spin down the membranes and membrane-bound proteins in a crude extract, centrifuge for 45,000 RPM at 4<sup>o</sup>C for 1 hour.  With our MLS-50 mushroom-shaped rotor, this is 163,000xg at r<sub>average</sub> (108,000xg at r<sub>minimum</sub>).
* To spin down the membranes and membrane-bound proteins in a crude extract, centrifuge for 45,000 RPM at 4<sup>o</sup>C for 1 hour.  With our MLS-50 mushroom-shaped rotor, this is 163,000xg at r<sub>average</sub> (108,000xg at r<sub>minimum</sub>).
== Important Info about Rotor & Tubes ==
* ''' Tubes should never be less than half filled or they will collapse.'''  - Tech support 5/2013 via [[User:Janet B. Matsen|Janet]]
** This is true for every tube material and size.
* You can wash and re-use tubes, however, deformed tubes will collapse.
** Be careful to wash them with compatible liquids.  See the manuals.
* There are seals inside the swinging buckets (tube holders) that seal the sample when you screw the bucket lid tight.  These are very important.  The ultracentrifuge uses a strong vacuum to keep the samples chilled.  If the seal fails, your sample will be sucked out of the swinging basket and end up in the vacuum.  There is no way to recover after this.
** Tech support said we should replace them yearly and have spares on hand.  I am skeptical, as this doesnt factor in how heavily the system is used.
== Parts & Info ==
* A fuller product spec sheet for the MLS-50: [[image:MLS-50 product sheet.jpg|thumb|center|MLS-50 product sheet]] from [https://www.beckmancoulter.com/wsrportal/bibliography?docname=BR-8101.pdf website PDF]
**

Revision as of 09:19, 4 June 2013

Back to Protocols

Our System

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 (< half full) 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).

Important Info about Rotor & Tubes

  • Tubes should never be less than half filled or they will collapse. - Tech support 5/2013 via Janet
    • This is true for every tube material and size.
  • You can wash and re-use tubes, however, deformed tubes will collapse.
    • Be careful to wash them with compatible liquids. See the manuals.
  • There are seals inside the swinging buckets (tube holders) that seal the sample when you screw the bucket lid tight. These are very important. The ultracentrifuge uses a strong vacuum to keep the samples chilled. If the seal fails, your sample will be sucked out of the swinging basket and end up in the vacuum. There is no way to recover after this.
    • Tech support said we should replace them yearly and have spares on hand. I am skeptical, as this doesnt factor in how heavily the system is used.

Parts & Info

  • A fuller product spec sheet for the MLS-50:
    MLS-50 product sheet
    from website PDF