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

Day 0 (Apr 6 Mon)
 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)  MeOH sensors also detects ethanol! :See reference #1 on the bottom of the page

                4/6/2009 (00:20)  Prepare small culture for inoculum (10mL)                 4/6/2009 (04:00)  Prepare small culture for inoculum (175mL)

           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                 4/6/2009 (18:00)  Fermentation 00:00 Inoculation 50 ml each using pump.                 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)
          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                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)                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: <font style=background-color:yellow>(10 rpm) F2: Induce with Galactose (55 mL) Start feeding: <font style=background-color:yellow>(10 rpm) F3: Induced with Galactose (130 mL, diluted x2.4) Start feeding: <font style=background-color:yellow>(1 rpm with 32 rpm pump)

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