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Great Bay Community College
320 Corporate Drive
Portsmouth, NH 03801
Title: New Brunswick BioFlo 3000 Bioreactor SOP
Approvals:
Preparer: ___Kari Britt_______________________________ Date _____15May09______
Reviewer: __Bob O'Brien_____________________________ Date _____15May09______
1. Purpose: Operation of the New Brunswick BioFlo 3000 Bioreactor.
2. Scope: Applies to growing yeast or bacteria in the New Brunswick BioFlo 3000 Bioreactor.
3. Responsibilities:
3.1. It is the responsibility of the course instructor /lab assistant to ensure that this SOP is
performed as directed and to update the procedure when necessary.
3.2. It is the responsibility of the students/technicians to follow the SOP as described and to
inform the instructor about any deviations or problems that may occur while performing
the procedure.
4. References:
4.1. New Brunswick BioFlo 3000 Bioreactor instrumentation manual
4.2. BioCommand Lite Software Manual
4.3. autoclave SOP
5. Definitions: N/A
6. Precautions: N/A
7. Materials:
7.1. pH 4.0 buffer standard
7.2. pH 7.0 buffer standard
7.3. laboratory tissues
7.4. silicone tubing
7.5. PharMed tubing
7.6. clamps
7.7. aluminum foil
7.8. air filters
7.9. glass wool
7.10. probe stand with clamps
7.11. glycerol
7.12. laboratory glassware detergent
7.13. deionized water
7.14. DO electrolyte
7.15. 3M potassium chloride (KCl) solution for pH probe storage
7.16. bleach
7.17. Optional: antifoaming agent (such as Antifoam A)
7.18. personal computer to run BioCommand Lite Software and Microsoft Excel
7.19. Bioreactor
7.19.1. pH probe
7.19.2. DO probe
7.19.3. RTD temperature probe
7.19.4. harvest tube
7.19.5. sparger tube
7.19.6. thermowell tube
Document Number: 1.16.1
Revision Number: 5
Effective Date: 20May09
Page 1 of 20
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Summary of Contents for Brunswick BioFlo 3000
- Page 1 Reviewer: __Bob O’Brien_____________________________ Date _____15May09______ 1. Purpose: Operation of the New Brunswick BioFlo 3000 Bioreactor. 2. Scope: Applies to growing yeast or bacteria in the New Brunswick BioFlo 3000 Bioreactor. 3. Responsibilities: 3.1. It is the responsibility of the course instructor /lab assistant to ensure that this SOP is performed as directed and to update the procedure when necessary.
- Page 2 Note: All fittings are hand-tightened except the DO probe adapter. 8.2.1. Attach the DO, pH, and temperature cables to their appropriate outlets on the front side of the cabinet of the BioFlo 3000. 8.2.2. If necessary, insert the plastic plugs into the holes in the glass vessel.
- Page 3 Effective Date: 20May09 Page 3 of 20 Title: New Brunswick BioFlo 3000 Bioreactor SOP 8.2.7. If no leaks are observed, remove the water from the vessel by lifting the vessel (still attached to the heat exchanger base) and pouring the water out into a sink or container.
- Page 4 8.7.7. Turn on the water. The water valve is located under the lab bench where the BioFlo 3000 is located. The pressure gauge should read between 15 and 20psi when the water is turned on.
- Page 5 Portsmouth, NH 03801 Effective Date: 20May09 Page 5 of 20 Title: New Brunswick BioFlo 3000 Bioreactor SOP 8.8.12. Immerse the pH and RTD temperature probes into a pH 4 standard buffer solution. Allow a few minutes for the electrode to equilibrate.
- Page 6 Portsmouth, NH 03801 Effective Date: 20May09 Page 6 of 20 Title: New Brunswick BioFlo 3000 Bioreactor SOP 8.12. Prepare for Operation 8.12.1. Remove the clamp from the tubing attached to the top outlet of the condenser. 8.12.2. Place the vessel and heat exchanger base on the platform in front of the cabinet so that it fits on the three steel plugs.
- Page 7 Effective Date: 20May09 Page 7 of 20 Title: New Brunswick BioFlo 3000 Bioreactor SOP 8.12.20. Set the control mode to P-I-D by selecting the box in the “Control” row of the “Temp 1” column. Press ALTER until “P-I-D” appears and then press the ENTER key.
- Page 8 8.14.1.2. Remove the protective cap from the top of the DO probe and connect the DO Cable to the DO probe. 8.14.1.3. If necessary, turn on the BioFlo 3000. 8.14.1.4. Allow the probe to sit for at least 30 minutes.
- Page 9 Effective Date: 20May09 Page 9 of 20 Title: New Brunswick BioFlo 3000 Bioreactor SOP 8.14.11. Allow ten to thirty minutes for the vessel to equilibrate. 8.14.12. Press the SCREEN key and then use the arrow keys to select the <Calibration>...
- Page 10 Effective Date: 20May09 Page 10 of 20 Title: New Brunswick BioFlo 3000 Bioreactor SOP 8.15.2.4. Set the gases mode to DO control by selecting the box in the “Mode” row (it is the only box available in the “Mode” row). Press the ALTER key until “DO”...
- Page 11 BioCommand Lite Software. 8.17.2. Check that the personal computer is connected to the BioFlo 3000 cabinet. If they are not connected, request that the lab manager or course instructor connect the computer to the cabinet before proceeding.
- Page 12 Portsmouth, NH 03801 Effective Date: 20May09 Page 12 of 20 Title: New Brunswick BioFlo 3000 Bioreactor SOP 8.17.4. From the desktop open BioCommand Lite by double clicking on the BCLite icon. 8.17.5. You can choose to use an existing recipe or create a new recipe. To use an existing recipe follow the procedure in step 8.17.6.
- Page 13 Portsmouth, NH 03801 Effective Date: 20May09 Page 13 of 20 Title: New Brunswick BioFlo 3000 Bioreactor SOP BT220 [semester] [year]. Record file name in the batch record, then click “OK”. 8.17.7.16. From the “Recipe[#].B1:Setup.[#]” window, click on the “Views” tab.
- Page 14 8.21.4. Select browse and choose the file used during the run. 8.21.5. Export the data to Microsoft Excel. 8.21.6. Turn all the control modes (Temp, pH, Agit, DO, Feed 1-5) on the Bioflo 3000 controller to off. 8.21.7. Turn off the BioFlo 3000.
- Page 15 8.22.25. Wipe down the BioFlo cabinet with 10% bleach solution. 9. Attachments: 9.1. Figure 1: BioFlo 3000 Vessel and Cabinet 9.2. Figure 2: Vessel Parts 9.3. Figure 3: Headplate, Underside 9.4. Figure 4: Assembled Headplate, Top Side 9.5.
- Page 16 320 Corporate Drive Revision Number: 5 Portsmouth, NH 03801 Effective Date: 20May09 Page 16 of 20 Title: New Brunswick BioFlo 3000 Bioreactor SOP Figure 1: BioFlo 3000 Vessel and Cabinet 1. Heat Exchanger Base 15. Probe Stand 2. Jacket Water Line Adaptors 16.
- Page 17 Document Number: 1.16.1 320 Corporate Drive Revision Number: 5 Portsmouth, NH 03801 Effective Date: 20May09 Page 17 of 20 Title: New Brunswick BioFlo 3000 Bioreactor SOP Figure 2: Vessel Parts 1. Small Thumb Screw 9. Baffle 2. Large Thumb Screw 10. Nut 3.
- Page 18 320 Corporate Drive Revision Number: 5 Portsmouth, NH 03801 Effective Date: 20May09 Page 18 of 20 Title: New Brunswick BioFlo 3000 Bioreactor SOP Figure 3: Headplate, Underside 1. Addition Port 8. Lower Impeller 2. Thermowell Port 9. DO Probe Port 3.
- Page 19 320 Corporate Drive Revision Number: 5 Portsmouth, NH 03801 Effective Date: 20May09 Page 19 of 20 Title: New Brunswick BioFlo 3000 Bioreactor SOP Figure 4: Assembled Headplate, Top Side 1. DO Probe 9. Agitator Cover 2. pH Probe 10. Sparger Tube 3.
- Page 20 320 Corporate Drive Revision Number: 5 Portsmouth, NH 03801 Effective Date: 20May09 Page 20 of 20 Title: New Brunswick BioFlo 3000 Bioreactor SOP Figure 5: BioFlo 3000 Side View 1. Gas and Air Console 2. Cabinet 3. Power Switch 4. Feed Pump Figure 6: BioFlo 3000 Controller 1.
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