User:Fermenter User/Notebook/MACG E4

{| width="800"
 * style="background-color: #cdde95;" align="center"|
 * style="background-color: #cdde95;" align="center"|




 * align="center" style="background-color: #e5edc8;" |


 * colspan="2" style="background-color: #F2F2F2;" align="right"|Customize your entry pages 
 * colspan="2"|
 * colspan="2"|
 * colspan="2"|

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 E4 (F2)

Day -1 (Feb 16 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 #6]            2/16/2009 (10:30)  Pour media 2/16/2009 (11:00) pH calibration 2/16/2009 (11:42) Antifoam added 2/16/2009 (12:36) Autoclave bioreactor 3 empty bottles/fermenter Carbon source, Inoculum, Feeding bottle 2/16/2009 (14:15) Start Temp Control 2/16/2009 (15:00) Add Carbon source using pump at 30°C 2/16/2009 (15:15) Start F2 software

Day 0 (Feb 17 Tue)
Sung-Hye Class 9:30-11:00

           2/17/2009 (7:35)   Connect MeOH sensor 2/17/2009 (7:50)  Calibrate dO2 2/17/2009 (7:50)  Set dO2   2/17/2009 (8:40) Fermentation 0:00 Inoculation 30 ml each using a pump.                2/17/2009 (18:00) Fermentation 8:00 F2: OD (18.9) Contamination? (N)Budding well, clumping.

Day 1 (Feb 18 Wed)
               2/18/2009 (9:00)  Fermentation 24:00 F2: OD (51.2) Contamination? (N) Still budding, no vacuoles.                2/18/2009 (13:00)  Fermentation 28:00 F2: OD (49.8) Contamination? (N)                2/18/2009 (17:00)  Fermentation 31:00 F2: OD (52.1) Contamination? (N)

Day 2 (Feb 19 Thu)
Sung-Hye Class 9:30 - 11:00

               2/19/2009 (08:00)  Fermentation 47:00 F2: OD (52.2) Contamination? (N) still budding, no vacuoles. Equivalent culture in flask OD 38.0                2/19/2009 (11:00)  Fermentation 50:00 F2: OD (54.0) Contamination? (N)

<Sung-Hye and Isis>  2/19/2009 (13:20) Fermentation 52:20 1uM Pepstatin added to F2 to limit activity of aspartyl proteases (Paper 8) F2: Connect Pump to 290 ml of Feeding media Induction 00:00 Watson pump: 9 rpm (10rpm = 12.8 ml/h) <Isis>               2/19/2009 (14:00)  Fermentation 53:00 F2: OD (55.4) Contamination? (N) Induction 01:00 <Isis>               2/19/2009 (18:30)  Fermentation 57:30 F2: OD (76.3) Contamination? (N) Induction 5:30 Vacuoles forming, actively budding <Isis>               2/19/2009 (23:50) Fermentation 62:50 F2: OD (68.08) Contamination? (N) Induction 10:50

Day 3 (Feb 20 Fri)
<Isis>               2/20/2009 (8:00) Fermentation 71:00 F2: OD (132.1) Contamination? (N)vacuoles approx. 4/cell Induction 19:00 <Isis>               2/20/2009 (09:00) Fermentation 71:00 F2: OD (132.4) Contamination? (N) Induction 20:00 <Sung-Hye and Isis>  2/20/2009 (10:00) Fermentation 72:00 F2: Harvest Induction 21:00