Centrifuge STANDARD OPERATING PROCEDURE

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STANDARD OPERATING PROCEDURE Centrifuge Model: Sorvall, RC-3B Plus Manufacturer: Kendro Laboratory Products Location: Fermentation Support, 1620 Food Sciences Building Publication Date: 01/23/2015 Iowa State University does not discriminate on the basis of race, color, age, ethnicity, religion, national origin, pregnancy, sexual orientation, gender identity, genetic information, sex, marital status, disability, or status as a U.S. veteran. Inquiries regarding non-discrimination policies may be directed to Office of Equal Opportunity, 3350 Beardshear Hall, 515 Morrill Road, Ames, Iowa 50011, Tel. 515 294-7612, email eooffice@iastate.edu.

Description and Uses The Sorvall Model RC-3B Plus is a large-capacity centrifuge for low speed (up to 6,000 rpm) work. The centrifuge operates over a temperature range of -20 C to +40 C and a time range of up to 35 min (plus a Hold setting that allows operation without a time limit). The centrifuge has a speed selection range of 0-6,000 rpm. At 5,000 rpm, the centrifuge has a maximum relative centrifugal force of 7277 x g. Power Specifications Motor: Spindle driven, Sorvall RC-3B+, SN 9904205 Voltage/Amperage: 200-240V/Single Phase/30A Speed Settings: 0-6,000 rpm @ 60 Hz. Temperature Range: -20 C to +40 C Time Settings: Up to 30 min @ 60 Hz; up to 35 min @ 50 Hz. For further information on specifications, general operation and installation requirements, see Chapters 1 and 2 in the Sorvall RC-3B Plus Operating Instructions manual (filed in the Fermentation Facility, 1621 Food Sciences Building). Potential Hazards and Safety Precautions Electric Shock/High Voltage (200V-240V) Make sure that the wall outlet receptacle is properly wired and grounded, and matches the instrument s power cord and plug. Make sure the area around the outlet, floor and your hands are completely dry when plugging or unplugging the electrical cord to/from the outlet. Fire Safety/Explosion Hazard Do not use the centrifuge with materials capable of developing flammable or explosive vapors. Note: No flammable liquids are to be centrifuged! Spindle-Driven Pinch Points/Possible Entanglement of Extremities, Hair, Jewelry or Clothing Use only Sorvall rotors and accessories designed for this centrifuge. Do not exceed maximum speeds for the rotors used. Never try to slow or stop a rotor by hand. Make certain the chamber door is locked and secure before operating the centrifuge. Never try to override the door-interlock system while the rotor is spinning. Make sure to secure long hair and any loose clothing or jewelry before operating the centrifuge. 2

Biohazard/Possible Illness or Physical Damage to Tissues and Organs Normal operation may involve use of pathogenic, toxic, radioactive, or otherwise hazardous materials. Such materials should not be used in the centrifuge unless all necessary safety precautions have been taken (see Operating Instructions manual). Note: Never use flammable solvents in a centrifuge! Be sure to wear all required personal protective equipment that is listed below. Required Personal Protective Equipment Lab Coat Hair Net (Tie Back Long Hair) Safety Glasses or Goggles Rubber, Latex or Disposable Gloves No Loose Fitting Clothing No Open-toed or Open-heeled Shoes Training Required Training *Denotes courses offered online Machine & Site-Specific Training Fire Safety & Extinguisher Training* Laboratory Safety: Core Concepts* Recommended Training for Frequent Users Electrical Safety & Lockout/Tagout Laboratory Safety: Spill Procedures 3

Operation 1. Be sure to acquire all required training, including sitespecific training before operating the centrifuge. 2. Chapter 3 of the Operating Instructions manual provides a more complete description of how to operate the centrifuge and its rotors. Be sure to read the manual before using the centrifuge to become familiar with its correct operation and individual component parts. 3. Be sure to wear all required personal protective equipment listed. 4. Press the power switch to the ON position. The power switch is located in the upper right corner of the front cabinet panel, just below the chamber door (see Figure 1). 5. To open the chamber door, pull up on the door opener. This handle is located on the right side of the chamber door (refer to Figure 1). 6. Remove the lid and safely install the H-6000A/HBB-6 Rotor in the chamber according to the directions given in the Rotor Instruction manual (on file in the Fermentation Facility, 1621 Food Sciences Building). This rotor has six swinging buckets that accepts one-liter centrifuge bottles (see Figure 2). Note: It is recommended that the rotor be pre-cooled before operation, either in the centrifuge chamber or in a refrigerator/cold room. 7. Fill the one-liter centrifuge bottles with samples to be centrifuged. Note: Equally balance the bottles in pairs, otherwise the rotor will vibrate and could potentially damage the centrifuge. Once balanced, insert each bottle of the balanced pair opposite one another in the rotor buckets. Only use bottles specifically designed for centrifugation! 8. Replace the lid and tighten it onto the rotor as per the directions given in the Rotor Instruction manual (on file in the Fermentation Facility, 1621 Food Sciences Building). After making sure the rotor (and lid) are secure, carefully close the chamber door. Emergency Stop ON/OFF Switch Figure 1: The Sorvall Model RC-3B Plus Centrifuge The H-6000A/HBB-6 Centrifugal Rotor 4

9. For the following instructions, refer to the Control Panel schematic below (Figure 3): a. Set the TEMPERATURE dial to the desired temperature. b. Turn the TIME dial to the desired run time or set to HOLD to allow operation without a time limit. c. Turn the SPEED dial to the desired RPM setting. Note: For the H-6000A/HBB-6 Rotor, the maximum allowable speed is 5000 rpm. d. If the braking function is needed to slow the rotor after a spin, turn the BRAKE dial to the desired setting. Control Panel for the Sorvall Model RC-3B Plus Centrifuge 10. Once the TEMP C indicator matches the setting on the TEMP SET dial, press the START switch. Then, the centrifuge will accelerate to the selected speed and run for the selected time. 11. When the time expires, the rotor will coast or brake to a stop. At this point, the DOOR light turns on to indicate that the latch interlock has released. Once released, open the door by pulling up and lifting the door handle. Note: NEVER try to bypass the chamber door s interlock system. 12. Loosen the lid from the rotor and remove it. Then, retrieve the centrifuged samples from the rotor. 13. Repeat the listed procedure if more centrifugation is needed. When all samples have been centrifuged, press the power switch to the OFF position. Record your use in the available centrifuge log book and initiate clean-up procedures. 5

Clean-up Procedures 1. Wash all centrifuge bottles and buckets with warm soapy water. 2. Rinse the centrifuge bottle and buckets with hot tap water, followed by a cool, distilled-water rinse. 3. If necessary, remove the rotor and wash/rinse as listed above in steps 1 and 2. 4. Allow the clean bottles, buckets, and rotor to dry by placing them into a drying cart or placing them upside down on a pad of clean paper towels. 5. If there was any spillage in or on the centrifuge, be sure to wipe all surfaces clean with an absorbent Teri towel. Then, wipe with a damp, but clean, Teri towel to rinse the surfaces. 6. Clean the surrounding work area. 7. Before leaving the work area, initiate check-out procedures by having the centrifuge and its accessories inspected by the Fermentation Facility manager. Machine Care and Maintenance Report all operational problems to the Fermentation Facility manager. Centrifuge must be properly cleaned and inspected after each use. All final inspections are performed by the Fermentation Facility manager. Spindle base should be properly lubricated on a regular basis by the Fermentation Facility manager or a trained service technician. Accessories This centrifuge comes equipped with the H-6000A/HBB-6 Rotor. Note: For this rotor, a maximum of 5000 RPM (RCF = 7,277 x g) must not be breached. 6