User:Fermenter User/Notebook/MACG F6

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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 F6

Day -1 (Apr 20 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

                4/20/2009 (10:20)  Prepare small culture for inoculum (4mL)

           4/20/2009 (11:00)  Pour media 4/20/2009 (11:00) pH calibration No Antifoam added 4/20/2009 (12:07) Autoclave bioreactor 3 empty bottles/fermenter Base, Carbon source+Inoculation, Feeding 4/20/2009 (13:30) Cooling & Start Temp Control 4/20/2009 (14:15) Add carbon source using pumps 4/20/2009 (14:30) Connect Base (400 ml) Prime the pumps 4/20/2009 (14:35) Connect MeOH sensor 4/20/2009 (14:35) Calibrate dO2 4/20/2009 (14:35) Set dO2 4/20/2009 (14:35) Wrap bioreactors with foil 4/20/2009 (14:37) Start F1 software 4/20/2009 (14:38) Start F2 software 4/20/2009 (14:39) Start F3 software 4/20/2009 (14:45) Start pH control

                4/20/2009 (15:00)  Prepare small culture for inoculum (10mL)                 4/20/2009 (19:15)  Prepare small culture for inoculum (175mL)

Day 0 (Apr 21 Tue)
               4/21/2009 (09:00)  Fermentation 00:00 Inoculation 50 ml each using pump.                4/21/2009 (09:00) Fermentation 00:00 F1: OD (0.16) Contamination? (N) F2: OD (0.16) Contamination? (N) F3: OD (0.16) Contamination? (N)                4/21/2009 (09:00) Fermentation 00:00 F1: 400 ml                              F2: 370 ml                              F3: 370 ml                    4/21/2009 (14:00) Fermentation 05:00 F1: OD (0.59) Contamination? (N) F2: OD (0.55) Contamination? (N) F3: OD (0.57) Contamination? (N)

Day 1 (Apr 22 Wed)
           4/22/2009 (07:30) Fermentation 12:30 F1: 320 ml                              F2: 290 ml                              F3: 300 ml                              Autoclaved meeding bottles <Ann>               4/22/2009 (09:00) Fermentation 24:00 F1: OD (7.7) Contamination? (N) F2: OD (7.5) Contamination? (N) F3: OD (7.6) Contamination? (N) <SungHye>           4/22/2009 (09:00) Fermentation 24:00 F1: Change Temp to 15C F2: Change Temp to 22C <Ann>               4/22/2009 (09:00) Fermentation 24:00  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) <Ann>               4/22/2009 (11:00) Fermentation 26:00 F1: OD (8.1) Contamination? (N) F1: Induction 02:00 F2: OD (7.4) Contamination? (N) F2: Induction 02:00 F3: OD (7.5) Contamination? (N) F3: Induction 02:00 <Ann>               4/22/2009 (13:00) Fermentation 28:00 F1: OD (9.1) Contamination? (N) F1: Induction 04:00 F2: OD (8.0) Contamination? (N) F2: Induction 04:00 F3: OD (7.9) Contamination? (N) F3: Induction 04:00 <Ann>               4/22/2009 (15:00) Fermentation 30:00 F1: OD (8.7) Contamination? (N) F1: Induction 06:00 F2: OD (9.0) Contamination? (N) F2: Induction 06:00 F3: OD (8.2) Contamination? (N) F3: Induction 06:00 <Ann>               4/22/2009 (17:00) Fermentation 32:00 F1: OD (8.8) Contamination? (N) F1: Induction 08:00 F2: OD (8.6) Contamination? (N) F2: Induction 08:00 F3: OD (8.4) Contamination? (N) F3: Induction 08:00 <Ann>               4/22/2009 (19:00) Fermentation 34:00 F1: OD (8.7) Contamination? (N) F1: Induction 10:00 F2: OD (8.5) Contamination? (N) F2: Induction 10:00 F3: OD (9.1) Contamination? (N) F3: Induction 10:00 <Ann>               4/22/2009 (21:00) Fermentation 36:00 F1: OD (11.4) Contamination? (N) F1: Induction 12:00 F2: OD (9.7) Contamination? (N) F2: Induction 12:00 F3: OD (8.8) Contamination? (N) F3: Induction 12:00

Day 2 (Apr 23 Thu)
<Ann>               4/23/2009 (00:00) Fermentation 39:00 F1: OD (8.8) Contamination? (N) F1: Induction 15:00 F2: OD (8.4) Contamination? (N) F2: Induction 15:00 F3: OD (7.8) Contamination? (N) F3: Induction 15:00 <Ann>               4/23/2009 (00:00) Fermentation 39:00 F1: OD (10.3) Contamination? (N) F1: Induction 15:00 F2: OD (9.1) Contamination? (N) F2: Induction 15:00 F3: OD (7.8) Contamination? (N) F3: Induction 15:00

Day 3 (Apr 24 Fri)
<Ann>               4/24/2009 (00:00) Fermentation 00:00 F1: OD (10.3) Contamination? (N) F1: Induction 15:00 F2: OD (9.1) Contamination? (N) F2: Induction 15:00 F3: OD (7.8) Contamination? (N) F3: Induction 15:00

<SungHye>           4/24/2009 (00:00) Harvest