User:Fermenter User/Notebook/MACG F4

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Background color codes:
Pastel Green: Major change of setup this time
yellow: Sung-Hye's correction or question to user

Letter color codes:
Black: Basic comment
Red: Emergency Information (Power shutdown, building access, Manager's absence etc)
Blue: Fermentation Setup information
Green: Discussion
Orange: Fermentation hrs
Purple: Induction hrs


MACG F4


Day -2 (Mar 22 Sun)

   <Ann>                 3/22/2009 (17:55)  Prepare small culture for inoculum (8mL)


Day -1 (Mar 23 Mon)

  <Bioreactor headplate configuation>
   pH sensor: monitor or control pH
   dO2 sensor: monitor or control dO2
   Condenser: Gas outlet
   OD probe: monitor cell growth ==> Not used! Replate it with septum
   Triple: Addition of media, Acid or Base if necessary
   MeOH sensor: Monitor EtOH (byproduct)
   
   <Sung-Hye's Note>
    MeOH sensors also detects ethanol! :See reference #1 on the bottom of the page
   


   <Sung-Hye>            3/23/2009 (11:30)  Pour media
                         3/23/2009 (11:30)  pH calibration
                                           No Antifoam added
                         3/23/2009 (12:48)  Autoclave bioreactor
                                            3 empty bottles/fermenter
                                            Base, Carbon source+Inoculation, Feeding

3/23/2009 (14:50) Start Temp Control 3/23/2009 (15:20) Wrap bioreactors with foil 3/23/2009 (15:21) Start F1 software 3/23/2009 (15:22) Start F2 software 3/23/2009 (15:24) Start F3 software 3/23/2009 (15:30) Connect MeOH sensor 3/23/2009 (15:38) Connect Base (400 ml) Prime the pumps 3/23/2009 (00:00) Add carbon source using pumps at 30°C


   <Ann>                 3/23/2009 (13:00)  Prepare small culture for inoculum (20mL)
   <Ann>                 3/23/2009 (18:50)  Prepare small culture for inoculum (220mL)

Day 0 (Mar 24 Tue)

   <Sung-Hye>           3/24/2009 (07:30)  Calibrate dO2
                        3/24/2009 (07:30)  Set dO2

<Sung-Hye and Ann> 3/24/2009 (07:30) Fermentation 00:00 Inoculation 50 ml each using pump. 3/24/2009 (07:30) Start pH control <Ann> 3/24/2009 (11:00) Fermentation 03:30 F1: OD (0.47) Contamination? (N) F2: OD (0.45) Contamination? (N) F3: OD (0.36) Contamination? (N) <Ann> 3/24/2009 (13:30) Fermentation 06:00 F1: OD (0.60) Contamination? (N) F2: OD (0.72) Contamination? (N) F3: OD (0.59) Contamination? (N) <Ann> 3/24/2009 (16:30) Fermentation 09:00 F1: OD (1.54) Contamination? (N) F2: OD (1.71) Contamination? (N) F3: OD (1.33) Contamination? (N)

Day 1 (Mar 25 Wed)

   <Ann>                3/25/2009 (08:00)  Fermentation 24:30 
                             F1: Induced with Galactose (55 mL)  F1: Induction 00:00  
                             Start feeding: (1 rpm) 
                             F2: Induced with Galactose (55 mL)  F2: Induction 00:00   
                             Start feeding: (10 rpm)    
                             F3: Induced with Galactose (55 mL)  F3: Induction 00:00 
                             Start feeding: (10 rpm) 
                             Spin down cells and re-suspend with fresh media
Flowate calibration of Watson pump for feeding. Click it to see large
Flowate calibration of Watson pump for feeding. Click it to see large
Flowate calibration of Watson pump (32 rpm) for feeding. Click it to see large
Flowate calibration of Watson pump (32 rpm) for feeding. Click it to see large


   <Ann>                3/25/2009 (08:00)  Fermentation 24:30 
                             F1: OD (7.76) Contamination? (N)F1: Induction 00:00 
                             F2: OD (7.38) Contamination? (N)F2: Induction 00:00 
                             F3: OD (6.08) Contamination? (N)F3: Induction 00:00 
   <Ann>                3/25/2009 (10:00)  Fermentation 26:30 
                             F1: OD (7.48) Contamination? (N)F1: Induction 02:00 
                             F2: OD (7.16) Contamination? (N)F2: Induction 02:00 
                             F3: OD (6.18) Contamination? (N)F3: Induction 02:00 
   <Ann>                3/25/2009 (12:00)  Fermentation 28:30 
                             F1: OD (7.92) Contamination? (N)F1: Induction 04:00 
                             F2: OD (7.38) Contamination? (N)F2: Induction 04:00 
                             F3: OD (6.24) Contamination? (N)F3: Induction 04:00 
   <Ann>                3/25/2009 (14:00)  Fermentation 30:30 
                             F1: OD (7.80) Contamination? (N)F1: Induction 06:00 
                             F2: OD (7.16) Contamination? (N)F2: Induction 06:00 
                             F3: OD (6.66) Contamination? (N)F3: Induction 06:00 
   <Ann>                3/25/2009 (16:00)  Fermentation 32:30 
                             F1: OD (8.02) Contamination? (N)F1: Induction 08:00 
                             F2: OD (7.00) Contamination? (N)F2: Induction 08:00 
                             F3: OD (7.02) Contamination? (N)F3: Induction 08:00 
   <Ann>                3/25/2009 (18:00)  Fermentation 34:30 
                             F1: OD (7.98) Contamination? (N)F1: Induction 10:00 
                             F2: OD (7.06) Contamination? (N)F2: Induction 10:00 
                             F3: OD (7.28) Contamination? (N)F3: Induction 10:00 
   <Ann>                3/25/2009 (21:00)  Fermentation 39:30
                             F1: Base consumption 400 ml - 300 ml = 100 ml total 
                             F2: Base consumption 400 ml - 300 ml = 100 ml total
                             F3: Base consumption 400 ml - 300 ml = 100 ml total
   <Ann>                3/25/2009 (21:00)  Fermentation 39:30 
                             F1: OD (9.00) Contamination? (N)F1: Induction 13:00 
                             F2: OD (8.60) Contamination? (N)F2: Induction 13:00 
                             F3: OD (8.50) Contamination? (N)F3: Induction 13:00 

Day 2 (Mar 26 Thu)

   <Ann>                3/26/2009 (00:00)  Fermentation 40:30 
                             F1: OD (9.40) Contamination? (N)F1: Induction 16:00 
                             F2: OD (8.60) Contamination? (N)F2: Induction 16:00 
                             F3: OD (8.50) Contamination? (N)F3: Induction 16:00 
   <Ann>                3/26/2009 (02:00)  Fermentation 42:30
                             F1: OD (10.5) Contamination? (N)F1: Induction 18:00 
                             F2: OD (8.80) Contamination? (N)F2: Induction 18:00 
                             F3: OD (10.8) Contamination? (N)F3: Induction 18:00 
   <Ann>                3/26/2009 (04:30)  Fermentation 45:00
                             F1: OD (10.3) Contamination? (N)F1: Induction 20:30 
                             F2: OD (8.90) Contamination? (N)F2: Induction 20:30 
                             F3: OD (9.40) Contamination? (N)F3: Induction 20:30 
   <Ann>                3/26/2009 (06:00)  Fermentation 46:30
                             F1: OD (8.70) Contamination? (N)F1: Induction 22:00 
                             F2: OD (9.50) Contamination? (N)F2: Induction 22:00 
                             F3: OD (9.10) Contamination? (N)F3: Induction 22:00 
   <Ann>                3/26/2009 (08:00)  Fermentation 48:30
                             F1: OD (9.10) Contamination? (N)F1: Induction 24:00 
                             F2: OD (8.40) Contamination? (N)F2: Induction 24:00 
                             F3: OD (9.50) Contamination? (N)F3: Induction 24:00 
   <Ann>                3/26/2009 (10:00)  Fermentation 50:30
                             F1: OD (9.30) Contamination? (N)F1: Induction 26:00 
                             F2: OD (8.90) Contamination? (N)F2: Induction 26:00 
                             F3: OD (8.90) Contamination? (N)F3: Induction 26:00 
   <Sung-Hye>           3/26/2009 (00:00)  Fermentation 00:00
                             F1: Base consumption (Not measured)
                             F2: Base consumption (Not measured)
                             F3: Base consumption (Not measured)
   <Ann>                3/26/2009 (10:00)  Fermentation 50:30
                             F1: Harvest F1: Induction 26:00
                             F2: Harvest F2: Induction 26:00 
                             F3: Harvest F3: Induction 26:00

References

  1. www.ravenbiotech.com/product.html [Paper1]


  1. High Cell Density Fermentation of Saccharomyces cerevisiae JUL3 in Fed-batch Culture for the Production of β-Glucan (2007) 13, 1, 153-158 [Paper2]
  Image:IE13-1-0153.pdf
  Fermentation condition: 30°C, 200 rpm, no pH control, 1 vvm
  
  1. Optimization of enterokinase fermentation using a recombinant Saccharomyces cerevisiae. (2005) 40, 717-722 [Paper3]
  Image:PB-40-717.pdf
  Fermentation condition: 30°C, 500 rpm, no pH control, 2 vvm
Error fetching PMID 16820461:
  1. Error fetching PMID 16820461: [Paper4]
  1. Peamiability test: Aliquots (100 µl) of this cell suspension were pipetted into fluoroplate wells, 
     which contained NPN (10 µM) and, as test substances, either EDTA (1.0 and 0.1 mM), PEI (10 µg ml–1), 
     DMSA (1 mM), AOT (1 mM), or HEPES buffer (control) to make up a total volume of 200 µl. If desired, 
     MgCl2 was added to the cell suspension before addition of NPN. Fluorescence was monitored within 
     3 min from four parallel wells per sample (excitation, 355 nm; half bandwidth, 38 ± 3 nm; emission, 
     402 nm; half bandwidth, 50 ± 5 nm). Each assay was performed at least three times. 
Error fetching PMID 9534237:
  1. Error fetching PMID 9534237: [Paper5]
  1. OD600 unit is equivalent to 0.64 +/- 0.02 g cells (dry weight)/L
Error fetching PMID 17111383:
  1. Error fetching PMID 17111383: [Paper6]
  1. To demonstrate the localization of the proteins relative to the cells, cell contours were visualized 
     with Alexa 647-concanavalin A.
Error fetching PMID 18451787:
  1. Error fetching PMID 18451787: [Paper7]
Error fetching PMID 17709746:
  1. Error fetching PMID 17709746: [Paper8]
Error fetching PMID 11005931:
  1. Error fetching PMID 11005931: [Paper8]

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