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Team:Hawaii/Notebook/2008-07-14
From 2008.igem.org
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(→Transformed DH5α with mutated GFP and new Biobrick vector constructs) |
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:* XbaI and PstI: GFP fusion brick and BBa_C0012 | :* XbaI and PstI: GFP fusion brick and BBa_C0012 | ||
::* Added XbaI and PstI separately | ::* Added XbaI and PstI separately | ||
| + | ::* Ran 25 μl of GFP and 20 μl of BBa_C0012 restriction digest reactions in gel | ||
:* HindIII and BamHI: Biobrick segment and pRL1383a | :* HindIII and BamHI: Biobrick segment and pRL1383a | ||
| + | ::* Ran 25 μl of Biobrick segment (half of total rxn) and 20 μl of pRL1383a restriction digest reactions in gel | ||
::* pRL1383a was not visbile under long wave UV. Need to rerun gel tomorrow with much more plasmid | ::* pRL1383a was not visbile under long wave UV. Need to rerun gel tomorrow with much more plasmid | ||
| Line 32: | Line 34: | ||
:<strong> Margaret</strong> | :<strong> Margaret</strong> | ||
| - | :*Transformed: pSB3K3, pSB1A2, pSB1AK3, (+)pUC18, (-)nothing | + | :*Transformed: pSB3K3, pSB1A2, pSB1AK3, (+)pUC18, (-)nothing. The positive and negative control verify that the DB3.1 competent cells made on 7/11/08 can take up DNA. The results of this test can be seen [[Team:Hawaii/Test Competent E. Coli|here]]. |
= Discussion = | = Discussion = | ||
= Quote of the Day = | = Quote of the Day = | ||
| - | <blockquote> | + | <blockquote></blockquote> |
{{Team:Hawaii/Footer}} | {{Team:Hawaii/Footer}} | ||
__NOTOC__ | __NOTOC__ | ||
Latest revision as of 01:23, 16 July 2008
| Projects | Events | Resources | ||
|---|---|---|---|---|
| Sponsors | Experiments | Milestones | Protocols | |
| Notebook (t) | Meetings (t) |
Things we did today
Wetlab work
PCR
- Grace
- 25 μl PCR reactions for GFP site directed mutagenesis and extraction of Biobrick pertinent sites from BBa_B0034
Made stocks
- Grace
- Made two 500mL batches of SOB
- Made 20 LB + amp100 plates
Transformed DH5α with ligations of a Biobrick vector with pnir, slr2016-1, slr2016-2, pilA, or C0012
- Grace
- Transformed using 5 μl of 1:1 and 1:10 ligation dilutions
- Transformed using 1 μl vector only (negative control)
- Incubated on ice 10 min as opposed to the recommended 30 min. by iGEM
RE digested PCR products and plasmids
- Grace
- XbaI and PstI: GFP fusion brick and BBa_C0012
- Added XbaI and PstI separately
- Ran 25 μl of GFP and 20 μl of BBa_C0012 restriction digest reactions in gel
- HindIII and BamHI: Biobrick segment and pRL1383a
- Ran 25 μl of Biobrick segment (half of total rxn) and 20 μl of pRL1383a restriction digest reactions in gel
- pRL1383a was not visbile under long wave UV. Need to rerun gel tomorrow with much more plasmid
Transformed DB3.1
- Margaret
- Transformed: pSB3K3, pSB1A2, pSB1AK3, (+)pUC18, (-)nothing. The positive and negative control verify that the DB3.1 competent cells made on 7/11/08 can take up DNA. The results of this test can be seen here.
Discussion
Quote of the Day
[http://manoa.hawaii.edu/ 
][http://manoa.hawaii.edu/ovcrge/ 
][http://www.ctahr.hawaii.edu 
]
