Team:Stanford/Notebook/Lab Work/Week 6
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*Performed first digestion (F2620 + sfGFP + pSB4k5) | *Performed first digestion (F2620 + sfGFP + pSB4k5) | ||
{| | {| | ||
- | | Part # | Part amount | BSA | NEB (#, amount) | Enzymes (1 uL each) | Water | + | | Part # || Part amount || BSA || NEB (#, amount) || Enzymes (1 uL each) || Water |
|- | |- | ||
- | | F2620 | 10.8 | 3 | 2, 3 | E + S | 11.2 | + | | F2620 || 10.8 || 3 || 2, 3 || E + S || 11.2 |
|- | |- | ||
- | | pSB4k5 | 20 | 3 | 3, 3 | E + P | 2 | + | | pSB4k5 || 20 || 3 || 3, 3 || E + P || 2 |
|- | |- | ||
- | | sfGFP | 11.76 | 3 | 2, 3 | X + P | 10.24 | + | | sfGFP || 11.76 || 3 || 2, 3 || X + P || 10.24 |
|} | |} | ||
Revision as of 17:42, 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 |