Team:Hawaii/Notebook/2008-09- 5
From 2008.igem.org
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Sponsors | Experiments | Milestones | Protocols | |
Notebook (t) | Meetings (t) |
Things we did today
Wetlab work
Construction of Broad-Host-Range Vector Intermediates
- Margaret
- Plasmid Prep: Everything that didn't work yesterday
- 2 Plac + B0030, 1 Plac + B0034, aadA(BB)colony 5, pSB3k3, pSB1A3
- Restriction Digest: same as Yesterday
- Ligation
- Transformation into DH5alpha (we need more competent cells)
Verification of transformants from yesterday
- Grace
Construct | Colonies |
---|---|
plac+rbs (no vector) | 0 |
C0012 derived vector | 12 |
C0012 derived vector, SAP treated | 163 |
3A: plac+rbs+vector | >300 |
- BBpRL1338a related plates all had a lawn (no distinct colonies observed). Too low antibiotic conc.? Will attempt to restreak/replate on Mon.
- Colony PCR of 3A assembly related colonies
- Oops. Loaded 1 μl colony #11 into colony #10's well
- Colony #10 has potential. Will restreak on Mon.
Drylab Work
Sequencing
- Margaret
- Analysis of Sequencing submitted (oriV1-4, 2 aadA(BB), 2 aadA (pRL))
- Grace
- Confirmed:
- nir+rbs
- rbs+GFP
- YAY!
Discussion
Quote of the Day
History is the only laboratory we have in which to test the consequences of thought. - Étienne Gilson
[http://manoa.hawaii.edu/ ][http://manoa.hawaii.edu/ovcrge/ ][http://www.ctahr.hawaii.edu ]