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Team:Hawaii/Notebook/2008-09- 5
From 2008.igem.org
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===Verification of transformants from yesterday=== | ===Verification of transformants from yesterday=== | ||
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[[Image:090508colonypcr.jpg|right|thumb|400px|EtBr stained 2% agarose gel ran at 83V for 1.75 hours. Two microliters of PCR product were loaded into each well. ]] | [[Image:090508colonypcr.jpg|right|thumb|400px|EtBr stained 2% agarose gel ran at 83V for 1.75 hours. Two microliters of PCR product were loaded into each well. ]] | ||
| + | :<strong>Grace</strong> | ||
| + | |||
{|class=wikitable border=1 align=center | {|class=wikitable border=1 align=center | ||
!Construct | !Construct | ||
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:* BBpRL1338a related plates all had a lawn (no distinct colonies observed). Too low antibiotic conc.? Will attempt to restreak/replate on Mon. | :* 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 | :* 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. | ::* Colony #10 has potential. Will restreak on Mon. | ||
| Line 42: | Line 44: | ||
:* Analysis of Sequencing submitted (oriV1-4, 2 aadA(BB), 2 aadA (pRL)) | :* Analysis of Sequencing submitted (oriV1-4, 2 aadA(BB), 2 aadA (pRL)) | ||
| + | :<strong>Grace</strong> | ||
| + | |||
| + | :* Confirmed: | ||
| + | ::* nir+rbs | ||
| + | ::* rbs+GFP | ||
| + | |||
| + | ::::<strong> YAY!</strong> | ||
= Discussion = | = Discussion = | ||
Latest revision as of 02:15, 6 September 2008
| Projects | Events | Resources | ||
|---|---|---|---|---|
| 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 
]
