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Team:Hawaii/Notebook/2008-08-22
From 2008.igem.org
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== Wetlab work == | == Wetlab work == | ||
===Verification of transformants=== | ===Verification of transformants=== | ||
| - | [[Image:082208colonypcr.jpg|right|thumb| | + | [[Image:082208colonypcr.jpg|right|thumb|400px|EtBr stained 2% agarose gel ran at 95V for 1.5 hours. Five microliters of PCR reaction were loaded into each lane.]] |
:<strong> Grace</strong> | :<strong> Grace</strong> | ||
{|class=wikitable border=1 align=center | {|class=wikitable border=1 align=center | ||
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:*ligation of rep+pSB1A3, P1lytic region+pSB1A3, aada(BB&pRL1383a)+pSB1A3 | :*ligation of rep+pSB1A3, P1lytic region+pSB1A3, aada(BB&pRL1383a)+pSB1A3 | ||
| - | === | + | ===Omega interposon=== |
| - | + | :<strong>Margaret</strong> | |
| - | + | :*streak on Sm&Sp plate to confirm insertion. | |
| - | + | ||
= Discussion = | = Discussion = | ||
Latest revision as of 13:41, 23 August 2008
| Projects | Events | Resources | ||
|---|---|---|---|---|
| Sponsors | Experiments | Milestones | Protocols | |
| Notebook (t) | Meetings (t) |
Things we did today
Wetlab work
Verification of transformants
- Grace
| Construct | Colony forming units |
|---|---|
| nir+rbs | 187 |
| plac+rbs | 0 |
| rbs+GFP | 1 |
| GFP+tt | 0 |
- Colony PCR of transformants
- Ran PCR reaction on 2% agarose gel
- Can't tell 15bp difference between GFP and rbs+GFP
- Restreaked single colony and inoculated for plasmid prep to confirm
- nir+rbs colonies #11-19 appear promising (larger than nir)
- Restreaked colony #19 and inoculated for plasmid prep to confirm
Plasmid prep of J33207
- Grace
Overnight RE digest
- Grace
- BB-pRL1383a and J33207 with XbaI and PstI
- pSB1A2 with EcoRI and PstI
- GFP, GFPf, nir with EcoRI and SpeI
Made LB+amp100 plates
- Grace
Ligation
Margaret
- ligation of rep+pSB1A3, P1lytic region+pSB1A3, aada(BB&pRL1383a)+pSB1A3
Omega interposon
- Margaret
- streak on Sm&Sp plate to confirm insertion.
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 
]
