User:Fermenter User/Notebook/BROM A7

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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
Pink: MeOH Induction hrs

BROM A7 (F4)

Contents

Day -6 (Jun 12 Fri):

<Raymond>              6/12/2009 (12:00)     Bring BMXY (powder) to fermenter room


Day -4 (Jun 14 Sun):

 Building closure 9am - 5pm 
 Power shut-down 10:30 am - 1:30 pm 


Day -3 (Jun 15 Mon):

<Raymond>              6/15/2009 (14:00)     Prepare and autoclave additives:
                               Try to make higher concentration, for instance
                               3. Potassium Phosphate:  ?? L
                               4. YNB: ?? L
                               5. Biotin: ?? L
                               6. Glycerol: ?? L
                           What is final volume of additives?  7.5 L
<Sung-Hye>             6/15/2009 (12:30)     Autoclave Acid/Base valves, Acid/Base/Antifoam bottles, Water
                       6/15/2009 (13:58)      Start Fermenter #4 software 
                       6/15/2009 (14:20)     Calculate amount of water = 50L - additives volume (7.5-L) = 42.5L
                       6/15/2009 (14:30)     Pour BMXY powder and water(42.5 L total)
                       6/15/2009 (14:30)     Add 10 ml of Antifoam
                       6/15/2009 (14:48)      Sterilization Routine 
                       6/15/2009 (15:40)      Set rpm at 300 
                       6/15/2009 (15:40)      Purge Air 50 L/min 

Day -2 (Jun 16 Tue):

<Sung-Hye>             6/16/2009 (10:00)     Cool down bioreactor at 4°C 
<Raymond>              6/16/2009 (00:00)   Take a colony from MD plate in 5 ml of MD.    
                                          Allow it to gorw at 30°C for 24 hrs.

Day -1 (Jun 17 Wed):

<Raymond>              6/17/2009 (00:00)   Suspend down cells and resuspend in 50 ml BMGY. 
                                          Allow it to gorw at 30°C for 24 hrs.
<Raymond>              6/17/2009 (00:00)   Suspend down cells and resuspend in 2 x 500 ml BMGY. 
                                          Allow it to gorw at 30°C for 24 hrs.

Day 0 (Jun 18 Thu):

<Raymond>              6/18/2009 (10:00)    Bring additives to fermenter room
<Sung-Hye>             6/18/2009 (10:00)      Add Adjutives using pump                 
                                            Picture was taken during transfer of media 

<Sung-Hye> 6/18/2009 (14:00) Connect Acid bottle Acid (2000 ml) 6/18/2009 (14:00) Calibrate dO2 probes 6/18/2009 (14:00) Start pH control
<Sung-Hye/Raymond>     6/18/2009 (14:45 - 15:40 ) 
                                           Inoculate cells  (O.D: 14.2 Volume: 1000 ml) using pump.
<Raymond>              6/18/2009 (15:30)     Fermentation 00:00  
                        No OD check

Day 1 (Jun 19 Fri): MeOH Induction Day 0

<Sung-Hye>             6/19/2009 (09:00)     Fermentation 17:30  
                         Connect Base
                         Acid (1850 ml) Base (1600 ml)
                         Autoclave Base bottle and MeOH bottole with connected tube
<Sung-Hye/Raymond>     6/19/2009 (11:00)  Fermentation 19:30  MeOH Induction 00:00 
                        (O.D:22)  Contamination? (No) Acid (1850-1800 ml=50 ml) Base (1200-1100 ml=100 ml)
                                         Connect MeOH pump
<Raymond>              6/19/2009 (17:00)  Fermentation 25:30  MeOH Induction 06:00 
                         Contamination? (No) Acid (1700 ml Base (1100 ml)
                                         Connect MeOH pump

Day 2 (Jun 20 Sat): MeOH Induction Day 1

<Raymond>              6/20/2009 (11:00)  Fermentation 19:30  MeOH Induction 24:00 
                       (O.D:24.29)  Contamination? (No)  Activity (0.154 /ul) Acid (1600 ml) Base (1100 ml) MeOH (1 ml)
                                         Feed MeOH: Tubing disconnected!!
<Sung-Hye>             6/20/2009 (14:00)  Fermentation 22:30  MeOH Induction 27:00 
                                          Autoclave new bottles and tubing for Base and MeOH   
                       6/20/2009 (15:30)  Fermentation 24:00  MeOH Induction 28:30 
                                          Connect Base
                                          Connect MeOH (1000 ml) and a pump for continuous feeding:  
                                          750 ml/24hr (= 3 times x 0.5%)
                                        = 31.25 ml/hr 
                                        = 0.52 ml/min
                                         Use 2rpm Max Watson pump at rpm 25
Flowate calibration of Watson pump (2 rpm) for feeding. Click it to see large
Flowate calibration of Watson pump (2 rpm) for feeding. Click it to see large

Day 3 (Jun 21 Sun): MeOH Induction Day 2

<Raymond>              6/21/2009 (1:30pm)  Fermentation 43:30  MeOH Induction 36:00 
                       (O.D:23.15 )  Contamination? (Yes)  Activity (0.118/ul) Acid ( 1500 ml) Base (1050 ml) MeOH (400 ml left, then topped up.

Day 4 (Jun 22 Mon): MeOH Induction Day 3

<Raymond>              6/22/2009 (2:00pm)  Fermentation 00:00 
                       (O.D:24.59 )  Contamination? (Yes)  Activity (0.5/5ul from old assay buffer, 1.40/5ul from new assay buffer) Acid (1500ml) Base (1050 ml) MeOH (400ml)

Day 5 (Jun 23 Tue): MeOH Induction Day 4

<Raymond>              6/23/2009 (00:00)    Fermentation 00:00  MeOH induction 00:00   
                       (O.D:?? )  Contamination? (Yes/No)  Activity (?? /ul) Acid (?? ml) Base (?? ml) MeOH (?? ml) 

Day 6 (Jun 24 Wed): MeOH Induction Day 5

<Raymond>              6/24/2009 (00:00)    Fermentation 00:00  MeOH induction 00:00   
                       (O.D:?? )  Contamination? (Yes/No)  Activity (?? /ul) Acid (?? ml) Base (?? ml) MeOH (?? ml) 



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