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Team:Hawaii/Notebook/2008-08- 5
From 2008.igem.org
| Projects | Events | Resources | ||
|---|---|---|---|---|
| Sponsors | Experiments | Milestones | Protocols | |
| Notebook (t) | Meetings (t) |
Things we did today
Wetlab work
Transformation
- Grace
- Transformed 50 μl DB3.1 competent cells with 5 μl ligation reactions of p+r or g+t
- Incubated on ice 17 minutes after adding DNA
PCR contamination test (cont. from 8/2 and 8/4)
- Grace
- Ran an EtBr stained 2.5% gel at 95V for 2 hours
- Why aren't I getting consistent results for any of the lanes?
Construction of p+r and g+t, otra vez
- Grace
- 10 μl PCR reactions for nir, slr1, slr2, pilA, J33207 (in case transformation doesn't work)
- RE digest of PCR products (no gel purification; used 3 μl PCR reactions)
- Digested nir and J33207 with EcoRI and SpeI
- Digested slr1, slr2, pilA with XbaI and PstI
- Ran 20 μl of digests on 2% agarose gel stained with EtBr for 1.5 hours
Drylab Work
Sequencing
- Grace
- Reviewed sequencing results from 7/21, emailed CORE Hawaii with concerns
- Mislabeled samples
- Poor quality reads
Discussion
- Looks like our TriDye 2-log ladder has gone bad (500bp band splits into two bands). Norman will aliquot out new ladder.
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 
]
