User:Fermenter User/Notebook/BROM A17

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BROM A17

Day -2 (Nov 14 Sun):
          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) Ortho-phosphoric acid is borrowed from Molday lab PTM1 Salt is from BROM-A16

Day -1 (Nov 15 Mon): MEDIA PREPARATION DAY
        11/15/2010 (14:00)    Assemble bioreactors: 1. Calibrate pH sensors 2. 5.0-L of Basal Salts: 3. 1-ml Antifoam         11/15/2010 (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                  11/15/2010 (14:30)   Autoclave                11/15/2010 (16:00)   Autoclave completed! <--Door stuck!                  11/15/2010 (11:50 - 12:00)                                Transfer cells to fresh 55-ml BMGY media                                Incubator in Fermentation Suite used

Day 0 (Nov 16 Tue): INOCULATION DAY
        11/16/2010 (9:45)   Set temperature to 28.5°C         11/16/2010 (9:45)   Purge Air 1L/min at 900 rpm         11/16/2010 (9:50)   Adjust pH to 5.0 with 140 ml NH4OH         11/16/2010 (9:50)     Start pH control F1: No Acid Base, 1M (400 ml)         11/16/2010 (9:51)     Start Fermenter #1 software 11/16/2010 (10:00)    Calibrate dO2 probes 11/16/2010 (10:00)    Strt dO2 control           11/16/2010 (12:30)     Check contamination of starter culture <Dennis&Sung-Hye> 11/16/2010 (13:00)     Fermentation 0:00 Flask 1 (from frozen stock) OD: 15 Inoculate 45 ml/bioreactor <Sung-Hye>        11/16/2010 (16:30)     Add 22 ml of PTM1 Salt/bioreactor

Day 1 (Nov 17 Wed):
<Dennis>          11/17/2010 (12:00)     Fermentation 23:00 F1 Base(400 ml) OD(1.9) Contamination(N) Pressure(N) Change Filter(N)

Day 2 (Nov 18 Thu)
<Dennis>          11/18/2010 (8:00)     Fermentation 44:00 F1 Base(400 ml) OD(8.1) Contamination(N) Pressure(N) Change Filter(N) <Dennis>          11/18/2010 (15:00)    Fermentation 51:00 F1 Base(400 ml) OD(10.75) Contamination(N) Pressure(N) Change Filter(N)

Day 3 (Nov 19 Fri): <font color=#FF00FF> MeOH Induction Day 1
<Sung-Hye>        11/19/2010 (09:00)    Fermentation 69:00 Base (0 ml) pH <5.0 (Adjust pH to 5.0 using conc. NH4OH                            Refill Base (+400 ml with 2M)                             Remove 500-ml media                             Change filter                              <Dennis>           11/19/2010 (12:00)     Fermentation 71:00                              F1: Base(?? ml) OD(51.25) Contamination(N) Pressure(N) Change Filter(N)      <Sung-Hye>         11/19/2010 (13:00)     Fermentation 73:00 <font color=#FF00FF> MeOH induction 00:00                              Calibrate MeOH sensor  6287 mV = 0.5%                             Start MeOH induction

Day 4 (Nov 20 Sat): <font color=#FF00FF> MeOH Induction Day 2
<Dennis>          11/20/2010 (14:00)     Fermentation 96:00 <font color=#FF00FF> MeOH induction 24:00 F1: Base(350 ml) MeOH(450 ml) OD(45.3) Contamination(N) Pressure(N) Change Filter(N) Activity assay (9.24 units/100ul), (2.16 mg/5-L)

Day 5 (Nov 21 Sun): <font color=#FF00FF> MeOH Induction Day 3
<Dennis>          11/21/2010 (14:00)     Fermentation 120:00 <font color=#FF00FF> MeOH induction 48:00 F1: Base(350 ml) MeOH(425 ml) OD(46.6) Contamination(N) Pressure(N) Change Filter(N) Activity assay (21.49 units/100ul), (5.03 mg/5-L)

Day 6 (Nov 22 Mon): <font color=#FF00FF> MeOH Induction Day 4
<Dennis>          11/22/2010 (14:00)     Fermentation 144:00 <font color=#FF00FF> MeOH induction 72:00 F1: Base(340 ml) MeOH(400 ml) OD(49.25) Contamination(N) Pressure(N) Change Filter(N) Activity assay (23.03 units/100ul), (5.39 mg/5-L)

Day 7 (Nov 23 Tue): <font color=#FF00FF> MeOH Induction Day 5
<Dennis>          11/23/2010 (14:00)     Fermentation 168:00 <font color=#FF00FF> MeOH induction 96:00 F1: Base(300 ml) MeOH(375 ml) OD(49.0) Contamination(N) Pressure(N) Change Filter(N) Activity assay (21 units/100ul), (4.94 mg/5-L) Harvest


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