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Team:Georgia State/Notebook
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Contents |
Notebook
July 7, 2010
Melissa, Alykhan
- Prepared tris HCl
July 15, 2010
Virginia, Angie, Alykhan, Joe
- Cut plasmid and ligated
- P. pastoris cells ready in -80°C
July 14, 2010
Virginia, Joe, Angie, Alykhan
- 10 glycerol stocks of 12E
- P. pasoris competent cells
- Started, ready at 2pm
- Plamid resuspension buffer made
- LB agar aliquots
- warm in water bath
- one 10mL tube for 1L agar
- extraction of 12E+9A plasmid parts: white tubes in freezer
July 13, 2010
Virginia
- 10 glycerol stocks of each
- P. pastoris
- 9A E. coli
- Inoculated 12E broth for plasmid extract and glycerol stocks
- Plate of P. pastoris for quality control
July 8, 2010
Angie, Kendra, Melissa, Nishedh, Alykhan
- Diluted pichia cells in a volume of 50mL to a OD600 = .186+.201
- Transform pars 12E and 12M into E. coli
July 7, 2010
Joe, Kendra, Angie
- Replated colonies from 9A RFP plate (3 plates).
- Colonies on broth 100 µL for 9A but no fluorescence observed
- 9A survivor colonies replated onto 10 µg/mL ampicillin plates and incubated at 30°C
- Mother plate in fridge
- Made 1000mL YPD
- Growing P. anomala in YPD broth for competent cells protocol
July 6, 2010
Dan
- Replated E. coli culture
Alykhan
- Transformation 9A, 8I (digested) & GFP.
July 2, 2010
Alykhan and Virginia
- DNA purification and ligation
- Replated E. coli culture
July 1, 2010
Joe
- Plated transformed cells containing either GFP (control) or iGEM part onto LB plates with 10µg/mL ampicillin (2 plates each, 10mL or 100mL)
- Also plated ≈100mL (remaining amount) of cells onto LB plates not containing ampicillin.
- Spread plates with hockey stick and placed in 37°C at 7:35.
