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Team:Hawaii/Notebook/2008-08- 6
From 2008.igem.org
| Projects | Events | Resources | ||
|---|---|---|---|---|
| Sponsors | Experiments | Milestones | Protocols | |
| Notebook (t) | Meetings (t) |
Things we did today
Wetlab work
Checked transformants from yesterday
- Grace
- Colony PCR'd transformants
- 30 cycles, anneal at 62C, extend for 1 min.
- Ran on EtBr stained 2.0% agarose gel at 95V for 1 hour
- None of the transformations were successful :o(
Prepared PCR'd nir and J33207 (from yesterday) for transformation/ligation
- Grace
- Ran PCR products on EtBr stained 2.0% agarose gel at 60V for 100 min.
- nir band at 330bp confirmed
- J33207 = 4 bands (1.5kb, 2.5kb, 3.2kb, 10kb), none of which are correct
- [http://www.partsregistry.org partsregistry] says J33207 DNA is inconsistent
- Extracted nir and 1.5kb J33207 band from gel
- RE digested in 50 μl rxns with EcoRI and SpeI in NEBuffer 2
- Larger rxn volume may improve digest efficiency
- Incubated at 37C for 2.5 hours
- RE digested 10 μl J33207 plasmid prep with EcoRI and SpeI in NEBuffer 2
- To confirm plasmid prep; if good, will use for ligation rxn
- Incubated at 37C for 2 hours
- Ran RE digests on EtBr stained 2.0% agarose gel at 72V for 100 min.
Made LB+amp100 plates
- Grace and Margaret
Drylab work
Sequencing
- Grace
- Reply from CORE Hawaii
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 
]
