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We have a Sorvoll RC-5B centrifuge that is useful for spinning large volumes in 0.5L bottles or medium-sized (a few mL) volumes very quickly
- Holds 8*50 mL, spins at 20,000 RPM/36,900 RCF
- BUT Katya (circa 2010) recommends using it at <= 18,000 RPM
- Can't be autoclaved.
- Actually, Amanda might have autoclaved them before.
- Can't write on body; can only write on cap. Sandy says stickers don't stick.
- Can't be autoclaved.
- BUT Katya (circa 2010) recommends using it at <= 7,500
- For bigger bottles. (0.5L)
- Holds eppendorf (1.5mL) tubes
- Spins at 9,000 RPM/13,700 RCF
- Requires 3 pieces for installation:
- (1) the base that you put the filled bottles in
- (2) a matching black metal lid
- (3) a metal rod with threads at one end and a knob on the other.
- Place the based down, screw the lid on counter clockwise and then use the pin to secure it. Try lifting to ensure you secured it correctly.
Explanation of RCF
- For reference, the benchtop 50mL centrifuges (e.g. ST-15R with a TX-400 swinging bucket rotor) we have spin at 4696 RCF.
- There are stickers with maximum speeds. These were set by Katya, a Lidstrom Lab manager, circa 2011. It is unknown what the motivation was for keeping the rotors below maximum speed.
- pre-chill the rotor you plan to use (overnight in fridge)
- turn on centrifuge and set it to the desired temperature range
- The dial on the left (under the temperature gauge) is really two knobs; one turns the lower bound blue stick and the other turns the upper bound red stick.
- The centrifuge will start cooling as soon as you turn it on.
- Install the rotor of your choice
- The small SS-34 rotor requires a T-shaped piece of metal with a screw end on it to install. Turn counter clockwise to screw it on. Use the same piece to unscrew it upon completion.
- The large GS-3 rotor has 3 pieces: the base that you put the filled bottles in, a lid, and a metal rod with threads at one end and a knob on the other. Place the based down, screw the lid on counter clockwise and then use the pin to secure it. Try lifting to ensure you secured it correctly.
- Fill bottles with the cells you want to centrifuge:
- To turn on, you have to have the black line in the temperature gauge panel between the blue and red lines. Then the start button glows and you can begin spinning.
- Note: the centrifuge is slow to speed up and slow to lose speed as well.
- Remove centrifuged cultures
- The machine slows down slowly.
- Leave the lid of the centrifuge open for a few hours until it warms up to room temperature and any condensation has evaporated. This will reduce odors.
Example protocol for SIP3-4:
Janet's SIP3-4 centrifugation:
- Chill to 4oC with SS-34 rotor inside
- Fill correct tubes
- Spin for ___ min at ___ RCF at ___oC
Example protocol for AM1:
Centrifugation Math & Basics
- Sigma summary:
- sedimentation rate is proportional to RFC, and proportional to the square of the particle diameter (cell diameter).