User:Fermenter User/Notebook/MACG F2

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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 F2

Day -2 (Feb 15 Sun)
                2/15/2009 (13:45) Prepare for small culture (15mL).

Day -1 (Feb 16 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            2/16/2009 (10:30)  Pour media 2/16/2009 (11:00) pH calibration 2/16/2009 (11:42) Antifoam added 2/16/2009 (12:36) Autoclave bioreactor 2 empty bottles/fermenter Carbon source+Inoculation 2/16/2009 (14:15) Start Temp Control 2/16/2009 (15:00) Add carbon source using pumps at 30°C 2/16/2009 (15:12) F1: Connect Base (400 ml) Start pH control 2/16/2009 (15:17) Start F1 software 2/16/2009 (15:18) Start F3 software

                2/16/2009 (15:45)  Prepare small culture for inoculum. (215mL)

Day 0 (Feb 17 Tue)
Sung-Hye: CHBE Class Auditing 9:30 - 11:00

          2/17/2009 (7:35)  Connect MeOH sensor 2/17/2009 (7:45) Connect Acid 2/17/2009 (7:50) Calibrate dO2 2/17/2009 (7:50) Set dO2   2/17/2009 (8:40)  Fermentation 00:00 Inoculation 50 ml each using pump.                2/17/2009 (10:00) Fermentation 01:20 F1: OD (0.280) Contamination? (N) F3: OD (0.335) Contamination? (N)                2/17/2009 (10:40) Fermentation 02:00 F1: OD (0.300) Contamination? (N) F3: OD (0.380) Contamination? (N)                2/17/2009 (11:15) Fermentation 02:35 F1: OD (0.310) Contamination? (N) F3: OD (0.370) Contamination? (N)                2/17/2009 (11:50) Fermentation 03:10 F1: OD (0.390) Contamination? (N) F3: OD (0.55) Contamination? (N) Induced with Galactose (55 mL) F3: Induction 00:00 <Ann>               2/17/2009 (12:50) Fermentation 04:10 F1: OD (0.390) Contamination? (N) F3: OD (0.470) Contamination? (N) F3: Induction 01:00 <Ann>               2/17/2009 (13:50) Fermentation 05:10 F1: OD (0.510) Contamination? (N) F3: OD (0.480) Contamination? (N) F3: Induction 02:00 <Ann>               2/17/2009 (15:50) Fermentation 07:10 F1: OD (0.590) Contamination? (N) F3: OD (0.500) Contamination? (N) F3: Induction 04:00 <Ann>               2/17/2009 (17:50) Fermentation 09:10 F3: OD (1.09) Contamination? (N) F3: Induction 06:00 <Ann>               2/17/2009 (18:50) Fermentation 10:10 F1: OD (0.950) Contamination? (N) <Ann>               2/17/2009 (19:50) Fermentation 11:10 F3: OD (1.780) Contamination? (N) F3: Induction 08:00

Day 1 (Feb 18 Wed)
<Ann>               2/18/2009 (03:50) Fermentation 19:10 F1: OD (5.74) Contamination? (N) F3: OD (6.65) Contamination? (N) F3: Induction 16:00 <Sung-Hye>          2/18/2009 (07:30) Fermentation 22:50 F1: Base consumption (400 ml - 340 ml = 60 ml) <Ann>               2/18/2009 (08:50)  Fermentation 24:00 F3: OD (6.70) Contamination? (N) Complete Fermentation F3: Induction 21:00 F1: OD (7.20) Contamination? (N) F1: Induction 00:00 Start feeding: (1 rpm) <Ann>               2/18/2009 (12:50)  Fermentation 25:00 F1: OD (6.58) Contamination? (N) F1: Induction 04:00 <Ann>               2/18/2009 (16:50)  Fermentation 29:00 F1: OD (8.12) Contamination? (N) F1: Induction 08:00

Day 2 (Feb 19 Thu)
<Ann>               2/19/2009 (00:50) Fermentation 37:00 F1: OD (8.64) Contamination? (N) F1: Induction 16:00 <Ann>               2/19/2009 (06:50) Fermentation 43:00 F1: OD (9.04) Contamination? (N) F1: Induction 22:00 <Sung-Hye>          2/19/2009 (09:30) Fermentation 45:00 Measure Base (400 ml - 270 ml= 130 ml total) <Ann>               2/19/2009 (13:30) Fermentation 49:40 F1: OD (10.36) Contamination? (N) F1: Induction 28:40 <Ann>               2/19/2009 (17:50) Fermentation 53:00 F1: OD (10.00) Contamination? (N) F1: Induction 33:00 <Ann>               2/19/2009 (23:50) Fermentation 58:00 F1: OD (10.72) Contamination? (N) F1: Induction 38:00

Day 3 (Feb 20 Fri)
<Ann>               2/20/2009 (07:20) Fermentation 65:30 F1: OD (11.75) Contamination? (N) F1: Induction 45:30 <Sung-Hye>          2/20/2009 (10:00) Fermentation 68:10 F1: Base (400 ml - 170 ml = 230 ml total) F1: Induction 48:10 <Ann>               2/20/2009 (13:50) Fermentation 72:10 F1: OD (10.95) Contamination? (N) F1: Induction 52:00 <Ann and Sung-Hye>  2/20/2009 (15:00) Fermentation 73:10 F1: Harvest F1: Induction 52:10