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Team:Hawaii/Notebook/2008-08- 2
From 2008.igem.org
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::* Old primer solution + new PCR water + new Taq = band at 300bp = primer solution was contaminated | ::* Old primer solution + new PCR water + new Taq = band at 300bp = primer solution was contaminated | ||
::* New primer solution + old PCR water + new Taq = bands at 270bp and 350bp = PCR water was contaminated | ::* New primer solution + old PCR water + new Taq = bands at 270bp and 350bp = PCR water was contaminated | ||
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| + | [[Image:080208 PCR nir GFPf gel.jpg|left|thumb|275px|EtBr stained 2.5% agarose gel ran at 60V for 2 hours. Five microliters of each PCR reaction was loaded into each well.]] | ||
| + | [[Image:080208 PCR gel extraction.jpg|left|thumb|275px|Gel after excision of nir-1 and GFPf-1 bands.]][[Image:080208 PCR contamination.jpg|left|thumb|275px|EtBr stained 4% agarose gel ran at 95V for 90 min. Ten microliters of each PCR reaction was loaded into each well.]] | ||
| + | <br><br> | ||
===Cryostocked=== | ===Cryostocked=== | ||
:<strong> Grace</strong> | :<strong> Grace</strong> | ||
:* nir-1, GFPf-1 | :* nir-1, GFPf-1 | ||
| + | ::* Threw out old (incorrect) cryostocks of nir and GFPf | ||
= Discussion = | = Discussion = | ||
Latest revision as of 17:59, 3 August 2008
| Projects | Events | Resources | ||
|---|---|---|---|---|
| Sponsors | Experiments | Milestones | Protocols | |
| Notebook (t) | Meetings (t) |
Things we did today
Wetlab work
PCR
- Grace
- Colony PCR of nir-1, nir-2, GFPf-1, GFPf-2
- Annealed at 62C; extended for 1 min.
- Used all new PCR water, primer solution, and Green Taq
- Cut out bands for nir-1, GFP-1 (correct size)
- PCR of PCR components to test for contamination
- New primer solution + new PCR water + old Taq = no bands
- Old primer solution + new PCR water + new Taq = band at 300bp = primer solution was contaminated
- New primer solution + old PCR water + new Taq = bands at 270bp and 350bp = PCR water was contaminated
Cryostocked
- Grace
- nir-1, GFPf-1
- Threw out old (incorrect) cryostocks of nir and GFPf
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 
]
