User:Fermenter User/Notebook/BROM A16

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BROM A16

Day -2 (Nov 8 Mon):
          11/8/2010 Thaw glycerol stock and grow in 5-ml MD media (prepared in summertime) Incubator in Fermentation Suite used         11/8/2010 (before go home) Preparing 5-L of Basal Medium (Defined media) <-- Phosphoric Acid used Preparing 1-L of PTM1 salt

Day -1 (Nov 9 Tue): MEDIA PREPARATION DAY
        11/9/2010 (14:00)    Assemble bioreactors: 1. Calibrate pH sensors 2. 5.0-L of Basal Salts <-- Phosphoric Acid used 3. 1-ml Antifoam         11/9/2010 (14:00)   Prepare bottles 1) no Acid need                                                 2) Base, 500-ml (tubing connected) x2                                                  3) MeOH, 1-L (tubing connected) x2                                                  4) Additives/Inoculum, 1 x 500-ml (tubing connected) x2                                                  5) Water, 2-L                  11/9/2010 (15:22)   Autoclave                11/9/2010 (17:00)   Autoclave completed!                11/9/2010 (17:00)   Set temperature to 28.5°C                 11/9/2010 (17:00)   Purge Air 1L/min at 900 rpm                 11/9/2010 (17:30)   Adjust pH to 5.0 with 140 ml NH4OH                   11/9/2010 (11:50 - 12:00)                                Transfer cells to 110-ml BMGY media                                Incubator in Fermentation Suite used

Day 0 (Nov 10 Wed): INOCULATION DAY
        11/10/2010 (10:18)     Start Fermenter #1 software 11/10/2010 (10:20)    Start Fermenter #2 software 11/10/2010 (11:00)    Calibrate dO2 probes 11/10/2010 (11:00)    Strt dO2 control           11/10/2010 (11:30)     Check contamination of starter culture         11/10/2010 (16:30)     Add 22 ml of PTM1 Salt/bioreactor <Dennis&Sung-Hye> 11/10/2010 (12:00)     Fermentation 0:00 Flask 1 (from frozen stock) OD: 15.3 Flask 2 (from streak on MD plate) OD: 27.8 Inoculate 45 ml/bioreactor <Sung-Hye>        11/10/2010 (12:00)     Start pH control F1: No Acid Base (400 ml) F2: No Acid Base (400 ml)

Day 1 (Nov 11 Thu):
<Dennis>        11/11/2010 (12:00)     Fermentation 24:00 F1 (Frozen Stock) Base(400 ml) OD(-) Contamination(N but strangely shaped translucent structures were found) Pressure(N) Change Filter(N) F2 (from Streak) Base(400 ml) OD(-) Contamination(N but strangely shaped translucent structures were found) Pressure(N) Change Filter(N)

Day 2 (Nov 12 Fri)
<Dennis>        11/12/2010 (12:00)     Fermentation 48:00 F1 Base(400 ml) OD(1.1) Contamination(N) Pressure(N) Change Filter(N) F2 Base(400 ml) OD(2.25) Contamination(N) Pressure(N) Change Filter(N)

Day 3 (Nov 13 Sat)
<Dennis>          11/13/2010 (12:00)     Fermentation 72:00 <font color=#FF00FF> MeOH induction 24:00 F1: Base(400 ml) OD(2.1) Contamination(N) Pressure(N) Change Filter(N) F2: Base(400 ml) OD(3) Contamination(N) Pressure(N) Change Filter(N) After 3 days, we decided to end the fermentation due to lack of yeast growth.


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