Team:Stanford/Notebook/Lab Work/Week 6
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***Heat inactivate at 65 C for 20 min | ***Heat inactivate at 65 C for 20 min | ||
*Performed first digestion (F2620 + sfGFP + pSB4k5) | *Performed first digestion (F2620 + sfGFP + pSB4k5) | ||
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| Part # || Part amount || BSA || NEB (#, amount) || Enzymes (1 uL each) || Water | | Part # || Part amount || BSA || NEB (#, amount) || Enzymes (1 uL each) || Water | ||
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Revision as of 17:44, 4 August 2010
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8/2 Monday
Greg's Notebook
- Inoculated freezer stocks of most of our parts with Alex
- Began planning for promoter characterization project:
- Going to do 3A ligations of the form: promoter + superfolded GFP + Kan plasmid
- Process:
- Digest parts with at least 2 ug DNA
- PCR cleanup
- Diagnostic gel (remember to save uncut DNA for control)
- Ligate in PCR tubes
- Heat inactivate at 65 C for 20 min
- Performed first digestion (F2620 + sfGFP + pSB4k5)
Part # | Part amount | BSA | NEB (#, amount) | Enzymes (1 uL each) | Water |
F2620 | 10.8 | 3 | 2, 3 | E + S | 11.2 |
pSB4k5 | 20 | 3 | 3, 3 | E + P | 2 |
sfGFP | 11.76 | 3 | 2, 3 | X + P | 10.24 |