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Team:Stanford/Notebook/Lab Work/Week 6
From 2010.igem.org
(Difference between revisions)
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== 8/2 Monday == | == 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 | ||
| + | |} | ||
Revision as of 17:41, 4 August 2010
| Home | Project | Applications | Modeling | Parts | Team | Notebook |
Spring: Brainstorming | Spring Meetings
Summer: Week 1 | Week 2 | Week 3 | Week 4 | Week 5 | Week 6 | Week 7 | Week 8 | Week 9 | Week 10 | Summaries
|
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 amount | BSA | NEB (#, amount) | Enzymes (1 uL each) | Water |
| 10.8 | 3 | 2, 3 | E + S | 11.2 |
| 20 | 3 | 3, 3 | E + P | 2 |
| 11.76 | 3 | 2, 3 | X + P | 10.24 |
