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Team:Hawaii/Notebook/2008-08-12
From 2008.igem.org
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::* plac+rbs with GFP, slr, pilA | ::* plac+rbs with GFP, slr, pilA | ||
:* Transformed into DH5α | :* Transformed into DH5α | ||
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| + | ===Rear ligation of signal sequences and GFPf=== | ||
| + | :<strong>Grace</strong> | ||
| + | |||
| + | :* Ligated slr and pilA with XbaI and PstI digested GFPf | ||
| + | :* Transformed into DH5&alpha using 5 μl ligation reaction | ||
== Drylab Work == | == Drylab Work == | ||
Revision as of 22:35, 12 August 2008
| Projects | Events | Resources | ||
|---|---|---|---|---|
| Sponsors | Experiments | Milestones | Protocols | |
| Notebook (t) | Meetings (t) |
Things we did today
Wetlab work
Test of lab's RE (cont.)
- Grace
- Ran RE digests from yesterday on a 1.2% agarose gel
- BamHI and HindIII digested DNA not visualized. Too little DNA added?
- NotI works. All plasmid preps except I14032 resulted in 2 bands of the correct size.
- Note: There is genomic DNA in all preps (band that looks like fire up top)
Verified transformants
- Grace
| Construct | Colony forming units |
|---|---|
| GFP+B0015(tt) | 1 |
| GFPf+B0015(tt) | 52 |
| slr1+GFPf | 56 |
| pilA+GFPf | 43 |
- Colony PCR of transformants to check for correct insert
- GFP+tt = unsuccessful
- GFPf+tt colonies 1, 3, 5 good
- slr1+GFPf colony 5 potentially good
- pilA+GFPf colonies 1, 2, 3, 4 potentially good
- Ligated slr, pilA with GFPf (ES digested) instead of GFPf (XP digested). Still got colonies, some which are potentially good. Hm...
- Restreaked:
- GFPf+tt colony 5
- slr1+GFPf colony 5
- pilA+GFPf colonies 1 and 4
- Will colony PCR again tomorrow to prep for sequencing
3A assembly
- Grace
- Ligated p+r with g or s onto pSB1A2
- pnir+rbs with GFP, slr1, pilA
- plac+rbs with GFP, slr, pilA
- Transformed into DH5α
Rear ligation of signal sequences and GFPf
- Grace
- Ligated slr and pilA with XbaI and PstI digested GFPf
- Transformed into DH5&alpha using 5 μl ligation reaction
Drylab Work
Name of Task
- name of person/people who performed the task
- Summary of task and what was done. Link to experiment for detailed notes if necessary.
- e.g. read through papers, worked on proposal, etc.
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 
]
