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

Day -1 (Mar 23 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

           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

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

Day 0 (Mar 24 Tue)
          3/24/2009 (07:30)  Calibrate dO2 3/24/2009 (07:30) Set dO2   3/24/2009 (07:30)  Fermentation 00:00 Inoculation 50 ml each using pump. 3/24/2009 (07:30) Start pH control                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)                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)                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: <font style=background-color:yellow>(1 rpm) F2: Induced with Galactose (55 mL) F2: Induction 00:00 Start feeding: <font style=background-color:yellow>(10 rpm) F3: Induced with Galactose (55 mL) F3: Induction 00:00 Start feeding: <font style=background-color:yellow>(10 rpm) <font style=background-color:yellow>Spin down cells and re-suspend with fresh media <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