User:Fermenter User/Notebook/BROM A18

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BROM A18

Day -2 (Nov 22 Mon):
          11/22/2010 Thaw glycerol stock and grow in 5-ml MD media (prepared in summertime) Incubator in Fermentation Suite used           11/22/2010 (before go home) Preparing 5-L of Basal Medium (Defined media) Ortho-phosphoric acid (85%) was ordered PTM1 Salt is form BROM-A16

Day -1 (Nov 23 Tue): MEDIA PREPARATION DAY
        11/23/2010 (14:00)    Assemble bioreactors: 1. Calibrate pH sensors 2. 5.0-L of Basal Salts (w/o Glycerol): 3. 1-ml Antifoam         11/23/2010 (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                  11/23/2010 (14:30)   Autoclave                11/23/2010 (16:00)   Autoclave completed!                  11/23/2010 (11:50 - 12:00)                                Transfer cells to fresh 55-ml BMGY media                                Incubator in Fermentation Suite used

Day 0 (Nov 24 Wed): INOCULATION DAY
        11/24/2010 (11:00)   Set temperature to 28.5°C         11/24/2010 (11:00)   Purge Air 1L/min at 900 rpm         11/24/2010 (11:30)     Add 22 ml of PTM1 Salt/bioreactor Add 158 ml of 50% Glycerol         11/24/2010 (11:00)   Adjust pH to 5.0 with 140 ml NH4OH         11/24/2010 (11:00)     Start pH control F1: No Acid Base, 1M (< 400 ml)         11/24/2010 (11:00)     Start Fermenter #1 software 11/24/2010 (11:00)    Calibrate dO2 probes 11/24/2010 (11:00)    Strt dO2 control <Dennis>          11/24/2010 (13:30)     Check contamination of starter culture <Dennis&Sung-Hye> 11/24/2010 (14:30)     Fermentation 0:00 Flask (from streak stock) OD: 10 Inoculate 45 ml/bioreactor

Day 1 (Nov 25 Thu):
<Dennis>          11/25/2010 (14:30)     Fermentation 24:00 F1 Base(300 ml) OD(11.3) Contamination(N) Pressure(N) Change Filter(N)

Day 2 (Nov 26 Fri) <font color=#FF00FF> MeOH Induction Day 1
<Dennis>          11/26/2010 (12:00)     Fermentation 46:00 F1 Base(210 ml) OD(17.25) Contamination(N) Pressure(N) Change Filter(N) <Sung-Hye>        11/26/2010 (12:30 - 15:30) Fermentation 48:00 F1 Fed-Batch <Dennis>          11/26/2010 (15:00)     Fermentation 48:00 <font color=#FF00FF> MeOH induction 00:00 F1 OD (19.05) <Sung-Hye>        11/26/2010 (15:30)     Fermentation 48:00 <font color=#FF00FF> MeOH induction 00:00 Calibrate MeOH sensor 6850 mV = 0.5% Start MeOH induction <Sung-Hye>        11/26/2010 (15:30)     Fermentation 48:00 <font color=#FF00FF> MeOH induction 00:00 Refill Base (150 ml + 250 ml = 400 ml)

Day 3 (Nov 27 Sat): <font color=#FF00FF> MeOH Induction Day 2
<Dennis>          11/27/2010 (14:00)     Fermentation 96:00 <font color=#FF00FF> MeOH induction 24:00 F1: Base(250 ml) MeOH(300 ml) OD(39.8) Contamination(N) Pressure(N) Change Filter(N) Activity assay (9.79 units/100ul), (2.29 mg/5-L)

Day 4 (Nov 28 Sun): <font color=#FF00FF> MeOH Induction Day 3
<Dennis>          11/28/2010 (14:00)     Fermentation 120:00 <font color=#FF00FF> MeOH induction 48:00 F1: Base(240 ml) MeOH(270 ml) OD(35.05) Contamination(N) Pressure(N) Change Filter(N) Activity assay (21.28 units/100ul), (4.98 mg/5-L)

Day 5 (Nov 29 Mon): <font color=#FF00FF> MeOH Induction Day 4
<Dennis>          11/29/2010 (14:00)     Fermentation 144:00 <font color=#FF00FF> MeOH induction 72:00 F1: Base(240 ml) MeOH(250 ml) OD(37.8) Contamination(N) Pressure(N) Change Filter(N) Activity assay (22.59 units/100ul), (5.29 mg/5-L)

Day 6 (Nov 30 Tue): <font color=#FF00FF> MeOH Induction Day 5
<Dennis>          11/30/2010 (8:30)     Fermentation 168:00 <font color=#FF00FF> MeOH induction 96:00 F1: Base(240 ml) MeOH(245 ml) OD(37.8) Contamination(N) Pressure(N) Change Filter(N) Activity assay (25.58 units/100ul), (5.99 mg/5-L) <Dennis>          11/30/2010 (11:30)     Fermentation 168:00 <font color=#FF00FF> MeOH induction 96:00 F1: Base(240 ml) MeOH(245 ml) OD(37.8) Contamination(N) Pressure(N) Change Filter(N) Activity assay (24.71 units/100ul), (5.78 mg/5-L) <Sung-Hye>        HARVEST


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