User:Fermenter User/Notebook/MACG F3

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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 F3

Day -1 (Mar 2 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

                3/2/2009 (09:30)  Prepare small culture for inoculum (5mL)            3/2/2009 (10:30)  Pour media 3/2/2009 (10:30) pH calibration No Antifoam added 3/2/2009 (11:11) Autoclave bioreactor 3 empty bottles/fermenter Base, Carbon source+Inoculation, Feeding 3/2/2009 (13:00) Wrap bioreactors with foil 3/2/2009 (13:34) Start Temp Control 3/2/2009 (14:00) Add carbon source using pumps at 30°C 3/2/2009 (14:05) Connect Base (400 ml) Prime the pumps 3/2/2009 (14:09) Start F1 software 3/2/2009 (14:13) Start F2 software 3/2/2009 (14:16) Start F3 software 3/2/2009 (16:00) Connect MeOH sensor

                3/2/2009 (14:05)  Prepare small culture for inoculum (10mL)                 3/2/2009 (18:40)  Prepare small culture for inoculum (175mL)

Day 0 (Mar 3 Tue)
Sung-Hye: CHBE Class Auditing 9:30 - 11:00

          3/3/2009 (07:20)  Calibrate dO2 3/3/2009 (07:20) Set dO2   3/3/2009 (09:00)  Fermentation 00:00 Inoculation 50 ml each using pump. 3/3/2009 (09:00) Start pH control                3/3/2009 (10:00) Fermentation 01:00 F1: OD (0.29) Contamination? (N) F2: OD (0.32) Contamination? (N) F3: OD (0.29) Contamination? (N)                3/3/2009 (11:00) Fermentation 02:00 F1: OD (0.54) Contamination? (N) F2: OD (0.58) Contamination? (N) F3: OD (0.56) Contamination? (N)                3/3/2009 (11:00) Fermentation 02:00 F1: Induced with Galactose (55 mL) F1: Induction 00:00 <Ann>               3/3/2009 (12:00) Fermentation 03:00 F1: OD (0.32) Contamination? (N) F1: Induction 01:00 <Ann>               3/3/2009 (13:00) Fermentation 04:00 F1: OD (0.49) Contamination? (N) F1: Induction 02:00 F2: OD (0.62) Contamination? (N) F3: OD (0.58) Contamination? (N) <Ann>               3/3/2009 (15:00) Fermentation 06:00 F1: OD (0.62) Contamination? (N) F1: Induction 04:00 F2: OD (0.97) Contamination? (N) F3: OD (0.78) Contamination? (N) <Ann>               3/3/2009 (17:00) Fermentation 08:00 F1: OD (1.20) Contamination? (N) F1: Induction 06:00 F2: OD (1.34) Contamination? (N) F3: OD (1.11) Contamination? (N) <Ann>               3/3/2009 (19:00) Fermentation 10:00 F1: OD (1.90) Contamination? (N) F1: Induction 08:00 <Ann>               3/3/2009 (21:00) Fermentation 12:00 F1: OD (2.74) Contamination? (N) F1: Induction 10:00 F2: OD (3.02) Contamination? (N) F3: OD (2.82) Contamination? (N)

Day 1 (Mar 4 Wed)
<Ann>               3/4/2009 (01:00) Fermentation 16:00 F1: OD (6.30) Contamination? (N) F1: Induction 14:00 F2: OD (5.02) Contamination? (N) F3: OD (4.42) Contamination? (N) <Ann>               3/4/2009 (7:00) Fermentation 22:00 F1: OD (8.40) Contamination? (N) F1: Induction 20:00 F2: OD (6.86) Contamination? (N) F3: OD (6.99) Contamination? (N) <Sung-Hye>          3/4/2009 (07:00) Fermentation 00:00 F1: Base consumption 400 ml - 300 ml = 100 ml total F2: Base consumption 400 ml - 320 ml = 80 ml total F3: Base consumption 400 ml - 340 ml = 60 ml total <Ann>               3/4/2009 (09:00)  Fermentation 24:00 F1: OD (9.35) Contamination? (N) F1: Induction 22:00 F2: OD (6.49) Contamination? (N) F3: OD (6.26) Contamination? (N) <Ann>               3/4/2009 (09:00) Fermentation 24:00 F2: Induced with Galactose (55 mL) F2: Induction 00:00 F3: Induced with Galactose (55 mL) F3: Induction 00:00 Start feeding: <font style=background-color:yellow>(10 rpm) <-- Supposed to be 1 rpm <Ann>               3/4/2009 (11:00) Fermentation 26:00 F1: OD (11.54) Contamination (N) Harvest F2: Induction 24:00 F2: OD (7.14) Contamination? (N) F2: Induction 02:00 F3: OD (7.68) Contamination? (N) F3: Induction 02:00 <Ann>               3/4/2009 (13:00) Fermentation 28:00 F2: OD (9.75) Contamination? (N) F2: Induction 04:00 F3: OD (10.55) Contamination? (N) F3: Induction 04:00 <Ann>               3/4/2009 (15:00) Fermentation 30:00 F2: OD (10.90) Contamination? (N) F2: Induction 06:00 F3: OD (11.70) Contamination? (N) F3: Induction 06:00 <Ann>               3/4/2009 (17:00) Fermentation 32:00 F2: OD (11.70) Contamination? (N) F2: Induction 08:00 F3: OD (12.55) Contamination? (N) F3: Induction 08:00 <Ann>               3/4/2009 (19:00) Fermentation 34:00 F2: OD (11.60) Contamination? (N) F2: Induction 10:00 F3: OD (13.50) Contamination? (N) F3: Induction 10:00 <Ann>               3/4/2009 (21:00) Fermentation 36:00 F2: OD (12.40) Contamination? (N) F2: Induction 12:00 F3: OD (12.50) Contamination? (N) F3: Induction 12:00 <Ann>               3/4/2009 (23:00) Fermentation 38:00 F2: OD (12.00) Contamination? (N) F2: Induction 14:00 F3: OD (13.60) Contamination? (N) F3: Induction 14:00

Day 2 (Mar 5 Thu)
<Ann>               3/5/2009 (01:00) Fermentation 40:00 F2: OD (12.40) Contamination? (N) F2: Induction 16:00 F3: OD (13.90) Contamination? (N) F3: Induction 16:00 <Ann>               3/5/2009 (03:00) Fermentation 42:00 F2: OD (13.10) Contamination? (N) F2: Induction 18:00 F3: OD (14.90) Contamination? (N) F3: Induction 18:00 <Ann>               3/5/2009 (05:00) Fermentation 44:00 F2: OD (12.00) Contamination? (N) F2: Induction 20:00 F3: OD (13.80) Contamination? (N) F3: Induction 20:00 <Ann>               3/5/2009 (07:00) Fermentation 46:00 F2: OD (12.80) Contamination? (N) F2: Induction 22:00 F3: OD (14.80) Contamination? (N) F3: Induction 22:00 <Sung-Hye>          3/5/2009 (07:00) Fermentation 00:00 F1: Base consumption 400 ml - 270 ml = 130 ml                             F2: Base consumption 400 ml - 220 ml = 180 ml                              F3: Base consumption 400 ml - 280 ml = 120 ml    <Ann>                3/5/2009 (09:00) Fermentation 48:00 F2: OD (12.90) Contamination? (N) F2: Induction 24:00 F3: OD (14.80) Contamination? (N) F3: Induction 24:00 <Ann>               3/5/2009 (11:00) Fermentation 50:00 F2: OD (12.20) Contamination? (Y/N) F2: Induction 26:00 F3: OD (15.20) Contamination? (Y/N) F3: Induction 26:00 <Ann>               3/5/2009 (11:00) Fermentation 50:00 F2: Harvest F2: Induction 26:00 F3: Harvest F3: Induction 26:00