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Team:Hawaii/Notebook/2008-08-25
From 2008.igem.org
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===Verification of transformants (cont.)=== | ===Verification of transformants (cont.)=== | ||
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[[Image:082508PCR.jpg|right|thumb|250px|EtBr stained 3% agarose gel ran at 60V for 3 hours. Two microliters of PCR reaction were loaded into each well.]] | [[Image:082508PCR.jpg|right|thumb|250px|EtBr stained 3% agarose gel ran at 60V for 3 hours. Two microliters of PCR reaction were loaded into each well.]] | ||
| + | :<strong> Grace</strong> | ||
| + | |||
:* PCR'd nir, nir+rbs, GFP, GFP+rbs | :* PCR'd nir, nir+rbs, GFP, GFP+rbs | ||
:* Ran on 3% agarose gel | :* Ran on 3% agarose gel | ||
| Line 25: | Line 26: | ||
::* C0012 with EcoRI and PstI (to isolate vector) | ::* C0012 with EcoRI and PstI (to isolate vector) | ||
::* nir with EcoRI and SpeI | ::* nir with EcoRI and SpeI | ||
| + | |||
| + | ===Ligation=== | ||
| + | :<strong>Krystle</strong> | ||
| + | |||
| + | :* Ligated overnight: | ||
| + | ::* plac + B0034 (both synthesized) | ||
| + | ::* plac + B0030 (both synthesized) | ||
| + | ::* gfpf (vector) + tt (synthesized) | ||
| + | ::* gfpf (re'd with E-S) + tt (synthesized) | ||
= Discussion = | = Discussion = | ||
Latest revision as of 03:05, 27 August 2008
| Projects | Events | Resources | ||
|---|---|---|---|---|
| Sponsors | Experiments | Milestones | Protocols | |
| Notebook (t) | Meetings (t) |
Things we did today
Wetlab work
Ligation & Transformation
- Margaret
- ligation of rep+pSB1A3 and P1+pSB1A3, transformation into DH5alpha cells
Colony PCR
Margaret
- verified the colonies from the (pLac+B0030) ligation and transformation. no transformants for the B0034 ligation
Verification of transformants (cont.)
- Grace
- PCR'd nir, nir+rbs, GFP, GFP+rbs
- Ran on 3% agarose gel
Restriction digest
- Grace
- RE digested overnight:
- BB-pRL1383a and J33207 with XbaI and PstI
- C0012 with EcoRI and PstI (to isolate vector)
- nir with EcoRI and SpeI
Ligation
- Krystle
- Ligated overnight:
- plac + B0034 (both synthesized)
- plac + B0030 (both synthesized)
- gfpf (vector) + tt (synthesized)
- gfpf (re'd with E-S) + tt (synthesized)
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 
]
