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Team:Hawaii/Notebook/2008-10- 9
From 2008.igem.org
| Projects | Events | Resources | ||
|---|---|---|---|---|
| Sponsors | Experiments | Milestones | Protocols | |
| Notebook (t) | Meetings (t) |
Things we did today
Wetlab work
Construction of secretion device (cont.)
- Grace
File:100908REdigest.jpg
EtBr stained 4% agarose gel ran at 60V for 3.5 hours. Twenty microliters of RE digested product were loaded into each well.
- Ran RE digests on gel
- Extracted bands from gel
- Ligated:
- J33207 and pRL1383a
- nir and rbs+GFPf+tt #1 and pSB1A3
- plac and rbs+GFPf+tt #1 and pSB1A3
- nir+rbs and slr1+GFPf and pSB1A3
- nir+rbs and pilA+GFPf+tt and pSB1A3
- Used 5 μl ligation reaction to transform DH5α
Sequencing
- Grace
- Prepared and sent samples in for sequencing:
- pilA+GFPf+tt #21 (KS)
- nir+rbs+slr1+GFPf #15 (10/2 transformation)
- slr1+GFPf+tt (10/2 transformation)
- nir+rbs+slr1+GFPf #6 (10/8 transformation)
- plac+rbs+pilA+GFPf+tt #5, 7, 11 (10/8 transformation)
Triparental conjugtion
- Grace
- BBpRL #1, 18 cultures did not grow
- PCR of colonies (on Xgal plate) showed no insert present (?!?)
- PCR of colonies from non-Xgal plate) showed no insert present (?!?)
- Faint band for colonies #1, 13 at 400bp ???
- Restreaked original colonies #1, 3, 13, 18 on freshly made LB+sp100+sm100
- Original colonies scraped w/ loop. Hopefully there's something there.
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 
]
