User:Fermenter User/Notebook/MOLD B1

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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
Purple: Induction hrs


MOLD B1


Day -2 (June 17 Tue):

      <Raj>              6/19/2010 (12:30)  
                                        Used BMGY media for first stage cell culturing 
                                        Prepared all necessary media components for fermentation:
                                         1. 0.5 L 10x YNB (filtered, excess amount)
                                         2. 0.5 L Phosphate buffer, pH 6.0 (autoclaved, excess amount)
                                         3. 0.5 L 10x Glycerol (filtered)
                                         4. 10mL 500x Biotin (filtered, excess amount)

Day -1 (June 16 Wed):

      <Raj>               6/19/2010 (10:30)     Leave the master culturing media containing the following items in the fermenter room:
                                                 1) Potassium Phosphate
                                                 2) YNB
                                                 3) Glycerol
<Raj> 6/19/2010 (10:30) Prepare 3-5L BMXY (1.4-L /fermenter x 2 + extra)
<Sung-Hye> 6/19/2010 (11:30) Assemble bioreactors: 1. Calibrate pH sensors 2. 2 x 1.4-L of BMXY: 3. 2 x 1-ml Antifoam
<Sung-Hye> 6/19/2010 (12:00) Prepared bottles: 1) Acid, 2 x 1-L (tubing separate) 2) Base, 2 x 500-ml (tubing connected) 3) MeOH, 2 x 1-L (tubing connected) 4) Additives/Inoculum, 2 x 1-L (tubing connected) 5) Water, 2-L

<Sung-Hye> 6/19/2010 (13:30) Autoclave Media leaked! (sample port was missing!) Sample ports tubing was connected and clamped! <Raj> 6/19/2010 (15:30) Made 2 x 1.4-L BMXY media again <Sung-Hye> 6/19/2010 (16:00) 1. 2 x 1.4-L of BMXY 2. 2 x 1-ml Antifoam 6/19/2010 (16:09) Autoclave again 6/19/2010 (17:45) Autoclave completed!

Day 0 (Jun 17 Thu):

      <Sung-Hye>          6/17/2010 (11:00)      Cooling bioreactors 
                                                 Purge Air 1 L/min at 100 rpm

Transfer Adjutives using pumps. 6/17/2010 (10:30-11:30) Combine: 3. 2 x 200 ml of 10x Potassium Phosphate 4. 2 x 200 ml of 10x YNB 5. 2 x 4 ml of 500x Biotin <--- from 4C 6. 2 x 200 ml of 10x Glycerol

6/17/2010 (13:30) Set rpm at 250 <-- little bit low 6/17/2010 (13:30) Calibrate dO2 probes 6/17/2010 (13:30) Fermenter #2: Start dO2 control (O2 valve Max) 6/17/2010 (13:45) Start pH control Acid (250 ml) Base (100 ml) <Raj&SungHye> 6/17/2010 (14:00) Fermentation 0:00 Flask OD: 1.5? Inoculate 2 fermentors (F#1 5-ml, F#2 3.5-ml) 6/17/2010 (15:16) Start Fermenter #1 software 6/17/2010 (15:18) Start Fermenter #2 software

Day 1 (Jun 18 Fri):

      <Raymond,SungHye,Raj>6/18/2010 (14:00)      Fermentation 24:00 
                                         Check contamination with Raymond.
                                         Both F1 and F2 were containing mainly bacteria, but rarely Pichia cells.
                                         Starter cultures were also containing heavily contaminated culture.
                                         Teminate fermentation.


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