User:Fermenter User/Notebook/MACG F5

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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 F5


Day -1 (Apr 5 Sun)

   <Ann>                 4/5/2009 (18:00)  Prepare small culture for inoculum (3mL)

Day 0 (Apr 6 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
   
   <Ann>                 4/6/2009 (00:20)  Prepare small culture for inoculum (10mL)
   <Ann>                 4/6/2009 (04:00)  Prepare small culture for inoculum (175mL)
   <Sung-Hye>            4/6/2009 (09:00)  Pour media
                         4/6/2009 (09:10)  pH calibration
                                           No Antifoam added
                         4/6/2009 (11:03)  Autoclave bioreactor
                                            3 empty bottles/fermenter
                                            Base, Carbon source+Inoculation, Feeding

4/6/2009 (13:20) Start Temp Control 4/6/2009 (13:20) Connect MeOH sensor 4/6/2009 (13:35) Wrap bioreactors with foil 4/6/2009 (14:24) Start F1 software 4/6/2009 (14:25) Start F2 software 4/6/2009 (14:26) Start F3 software 4/6/2009 (14:40) Connect Base (400 ml) Prime the pumps 4/6/2009 (14:50) Add carbon source using pumps at 30°C 4/6/2009 (14:52) Start pH control 4/6/2009 (14:55) Calibrate dO2 4/6/2009 (14:55) Set dO2

<Ann> 4/6/2009 (18:00) Fermentation 00:00 Inoculation 50 ml each using pump. <Ann> 4/6/2009 (18:00) Fermentation 00:00 F1: OD (0.20) Contamination? (N) F2: OD (0.13) Contamination? (N) F3: OD (0.12) Contamination? (N)

Day 1 (Apr 7 Tue)

   <Sung-Hye>           4/7/2009 (07:00)  Fermentation 13:00  
                             F1: 360 ml 
                             F2: 360 ml
                             F3: 330 ml
                        4/7/2009 (07:00)  Fermentation 13:00  
                             Change P-gain for F1 to 100
   <Ann>                4/7/2009 (11:30)  Fermentation 17:30 
                             F1: OD (6.90) Contamination? (N)
                             F2: OD (6.00) Contamination? (N)
                             F3: OD (3.60) Contamination? (N)
   <Ann>                4/7/2009 (18:00)  Fermentation 22:00 
                             F1: OD (9.10) Contamination? (N)
                             F2: OD (8.90) Contamination? (N)
                             F3: OD (6.00) Contamination? (N)
   <Ann>                4/7/2009 (18:00 - 20:30)  Fermentation 24:00
                             F1: Spin down cells and re-suspend with fresh media (1-L)
                                 Pour them into a pump and start pump 
                             F2: Spin down cells and re-suspend with fresh media (1-L)
                                 Pour them into a pump and start pump 
                             F3: Spin down cells and re-suspend with fresh media (1-L)
                                 Pour them into a pump and start pump 
   <Ann>                4/7/2009 (20:30)  Fermentation 26:30 
                             F1: OD (7.80) Contamination? (N)
                             F2: OD (7.70) Contamination? (N)
                             F3: OD (5.30) Contamination? (N)
   <Ann>                4/7/2009 (22:00)  Fermentation 28:00 
                             F1: OD (8.40) Contamination? (N)
                             F2: OD (8.50) Contamination? (N)
                             F3: OD (5.90) Contamination? (N)
   <Ann>                4/7/2009 (22:00)  Fermentation 28:00 Induction 00:00 
                             F1: Induce with Galactose (55 mL)  
                                 Start feeding: (10 rpm)
                             F2: Induce with Galactose (55 mL)                                     
                                 Start feeding: (10 rpm)
                             F3: Induced with Galactose (130 mL, diluted x2.4)  
                                 Start feeding: (1 rpm with 32 rpm pump)                              
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


Day 2 (Apr 8 Wed)

   <Ann>                4/8/2009 (04:00)  Fermentation 34:00 
                             F1: OD (11.60) Contamination? (N)F1: Induction 06:00 
                             F2: OD (11.20) Contamination? (N)F2: Induction 06:00 
                             F3: OD (10.90) Contamination? (N)F3: Induction 06:00 
   <Ann>                4/8/2009 (06:00)  Fermentation 36:00 
                             F1: OD (12.60) Contamination? (N)F1: Induction 08:00 
                             F2: OD (12.30) Contamination? (N)F2: Induction 08:00 
                             F3: OD (8.70) Contamination? (N)F3: Induction 08:00 
   <Ann>                4/8/2009 (08:00) HARVEST  Fermentation 38:00 
                             F1: OD (13.40) Contamination? (N)F1: Induction 10:00 
                             F2: OD (12.50) Contamination? (N)F2: Induction 10:00 
                             F3: OD (9.80) Contamination? (N)F3: Induction 10:00 
   <Sung-Hye>           4/8/2009 (08:00)  Fermentation 13:00  
                             F1: 280 ml 
                             F2: 180 ml
                             F3: 150 ml

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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