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Team:Hawaii/Notebook/2008-07-30
From 2008.igem.org
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=== Restriction Digest=== | === Restriction Digest=== | ||
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| - | + | [[Image:073008REdigests.jpg|EtBr stained 1.2% agarose gel. Twenty microliters of the restriction digest reactions were loaded into each well. Gel ran at 59V for 1.5 hours.]] | |
:* Digested 5 μl GFP, GFPf, and nir PCR products with EcoRI and SpeI | :* Digested 5 μl GFP, GFPf, and nir PCR products with EcoRI and SpeI | ||
:* Sequentially digested 10 μl pRL1383a and 5 μl J33207 PCR product with BamHI and HindIII | :* Sequentially digested 10 μl pRL1383a and 5 μl J33207 PCR product with BamHI and HindIII | ||
Revision as of 09:30, 31 July 2008
| Projects | Events | Resources | ||
|---|---|---|---|---|
| Sponsors | Experiments | Milestones | Protocols | |
| Notebook (t) | Meetings (t) |
Things we did today
Wetlab work
Gel Purification
- Grace
- Purified B0030 and B0024 from gel
- B0030 = 21.9 ng/μl
- B0024 = 5.7 ng/μl
Restriction Digest
- Grace
- Digested 5 μl GFP, GFPf, and nir PCR products with EcoRI and SpeI
- Sequentially digested 10 μl pRL1383a and 5 μl J33207 PCR product with BamHI and HindIII
- Used Roche enzymes and Roche buffer M (buffer B also works) from SC
- Ran digests on gel
- pRL1383a and J33207 bands good
- GFP, GFPf, and nir bands still too big
PCR
- Grace
- Redid yesterday's PCR of GFP, GFPf, and nir
PCR amplification of pRL1383a
- Margaret
- ran a gel from yesterday's PCR amplification of oriV, aadA, and rep regions.
Mini-Prep
- Margaret
- Plasmid Prep of [http://partsregistry.org/Part:BBa_I51020 BBa_I51020],[http://partsregistry.org/Part:pSB1A7 pSB1A7], [http://partsregistry.org/Part:BBa_B0015 BBa_B0015]
Colony PCR
- Margaret
- verify the insert of the oriT construct and [http://partsregistry.org/Part:BBa_I14032 BBa_I14032]
- Results:
- [http://partsregistry.org/Part:BBa_I14032 BBa_I14032] should be 334bp, the band under 0.4kb indicating a verification of this size of insert.
- oriT should be about 396bp, the band is around 0.4kb so is in approximately the right place.
- Discussion: The a plasmid prep and glycerol stock of both will be made. I need to verify the oriT part through sequencing.
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
The gel purification kit is almost out of PE. If we need more before our order comes in, there is PE in the Qiaprep Spin Miniprep kit (just remember to add EtOH before use).
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 
]
