User:Fermenter User/Notebook/BROM A7

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<!-- sibboleth --><div id="lncal1" style="border:0px;"><div style="display:none;" id="id">lncal1</div><div style="display:none;" id="dtext"></div><div style="display:none;" id="page">User:Fermenter User/Notebook/BROM A7</div><div style="display:none;" id="fmt">yyyy/MM/dd</div><div style="display:none;" id="css">OWWNB</div><div style="display:none;" id="month"></div><div style="display:none;" id="year"></div><div style="display:none;" id="readonly">Y</div></div>

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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)

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

File:Watsonpump.tif

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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