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Team:Hawaii/Notebook/2008-08- 7
From 2008.igem.org
| Projects | Events | Resources | ||
|---|---|---|---|---|
| Sponsors | Experiments | Milestones | Protocols | |
| Notebook (t) | Meetings (t) |
Things we did today
Wetlab work
Plasmid prep
- Krystle
- Finished plasmid prep of BB-pRL1383a, nir, GFPf from yesterday
Cyanobacteria seed stocks
- Krystle
- Reinoculated liquid and solid media seed stocks
Verification of transformants
- Grace
| Construct | Colony forming units |
|---|---|
| nir + B0030 (rbs) | 2 + 1 cluster of colonies |
| plac (I14032) + B0030 (rbs) | 37 |
| GFP (E0040) + B0015 (tt) | 8 |
| GFPf + B0015 (tt) | 4 |
| lac device (J33207) + B0015 (tt) | 56 |
- Colony PCR of transformants
- Ran on EtBr stained 2.5% agarose gel at 95V for 75 min.
- Looks like nir+rbs and I14032+rbs worked. YAY :o). Will prep for sequencing tomorrow.
- GFP, GFPf, J33207 don't look like they went in. Will sequence to confirm anyway.
Plasmid Prep
Margaret
- Re-did yesterday's plasmid prep of pSMC121 because I don't believe the band was big enough. I e-mailed Dr. Callahan to get the exact size, but I know, because it contains the omega interposon, that it is at least 2kb.
Extraction from Gel
Margaret
- extracted 3 bands of 50ul reaction for rep and oriV first by cutting out using long wave UV for visualization.
- Purification from gel with Qiagen Minelute gel purification kit.
PCR
Margaret
- Large-scale PCR amplification of aadA (it was lost in a gel cutting mishap) and p1 lytic.
Discussion
Quote of the Day
Gernot: "So would you eat the E. coli to save them if we lose the -80C in like a war?"
Loren: "Well, I'll eat them, but you'll be collecting them."
