User:Fermenter User/Notebook/MACG F7

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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 F7R

Day -1 (May 12 Tue)
 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

                5/12/2009 (9:10)  Prepare small culture for inoculum (10mL)

       5/12/2009 (11:30)  Pour media 5/12/2009 (11:30) pH calibration No Antifoam added 5/12/2009 (12:45) Autoclave bioreactor 3 empty bottles/fermenter Base, Carbon source+Inoculation, Feeding 5/12/2009 (14:30) Cooling & Start Temp Control 5/12/2009 (14:50) Add carbon source using pumps 5/12/2009 (15:05) Connect Base (400 ml) Prime the pumps Set pH                          5/12/2009 (15:10)  Start F1 software 5/12/2009 (15:11) Start F2 software 5/12/2009 (15:12) Start F3 software

                5/12/2009 (12:45)  Prepare small culture for inoculum (20mL)                 5/12/2009 (20:00)  Prepare small culture for inoculum (420mL)

Day 0 (May 13 Wed)
           5/13/2009 (08:40)  Connect MeOH sensor 5/13/2009 (08:40) Wrap bioreactors with foil 5/13/2009 (08:40) Calibrate dO2 5/13/2009 (08:40) Set dO2

               5/13/2009 (09:00)  Fermentation 00:00 Inoculation 50 ml (F1 and F2) and 250 ml (F3) using pump.                5/13/2009 (09:00) Fermentation 00:00 F1: OD (0.150) Contamination? (N) F2: OD (0.135) Contamination? (N) F3: OD (0.209) Contamination? (N)                5/13/2009 (10:00) Fermentation 01:00 F1: 380 ml                              F2: 380 ml                              F3: 360 ml                    5/13/2009 (14:00) Fermentation 05:00 F1: OD (0.633) Contamination? (N) F2: OD (0.635) Contamination? (N) F3: OD (0.610) Contamination? (N) <Ann>               5/13/2009 (17:30) Fermentation 08:30 F1: OD (1.086) Contamination? (N) F2: OD (1.123) Contamination? (N) F3: OD (1.089) Contamination? (N)

Expected Power Outrage 7:00 pm - 10:00 pm                  UPS (Undisrupted power supply) will be running

Day 1 (May 14 Thu)
<Ann>               5/14/2009 (07:00-08:00) Fermentation 22:00-23:00 F1: Spin down cells and re-suspend with fresh media (1-L) Pour culture back into fermenter <Ann>               5/14/2009 (09:00) Fermentation 24:00 F1: 290 ml                              F2: 300 ml                              F3: 30 ml (replaced with 400 mL base at 10x concentration) <Ann>               5/14/2009 (09:00) Fermentation 24:00 F1: OD (7.7) Contamination? (N) F2: OD (10.5) Contamination? (N) F3: OD (8.9) Contamination? (N) <Ann>               5/14/2009 (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 (275 mL) Start feeding: <font style=background-color:yellow>(2 rpm with 32 rpm pump) <Ann>               5/14/2009 (10:00) Fermentation 25:00 F3: 500 mL was taken out to avoid overflow / clogging the filter F3: Induction 01:00 <Ann>               5/14/2009 (11:00) Fermentation 26:00 F1: OD (8.8) Contamination? (N) F1: Induction 02:00 F2: OD (9.5) Contamination? (N) F2: Induction 02:00 F3: OD (8.4) Contamination? (N) F3: Induction 02:00 <Ann>               5/14/2009 (13:00) Fermentation 28:00 F1: OD (9.8) Contamination? (N) F1: Induction 04:00 F2: OD (9.0) Contamination? (N) F2: Induction 04:00 F3: OD (8.4) Contamination? (N) F3: Induction 04:00 <Ann>               5/14/2009 (15:00) Fermentation 30:00 F1: OD (10.7) Contamination? (N) F1: Induction 06:00 F2: OD (9.4) Contamination? (N) F2: Induction 06:00 F3: OD (8.7) Contamination? (N) F3: Induction 06:00 <Ann>               5/14/2009 (17:00) Fermentation 32:00 F1: OD (15.6) Contamination? (N) F1: Induction 08:00 F2: OD (11.0) Contamination? (N) F2: Induction 08:00 F3: OD (8.9) Contamination? (N) F3: Induction 08:00 <SungHye>           4/24/2009 (00:00) Harvest