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Team:Hawaii/Notebook/2008-07-17
From 2008.igem.org
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===PCR=== | ===PCR=== | ||
:<strong> Grace </strong> | :<strong> Grace </strong> | ||
| + | [[Image:071608 PCR.jpg|thumb|right|250px|PCR products on an EtBr stained 4% agarose gel run at 119V for 27 min then 95V for 25 min. Ten microliters of PCR reactions were loaded into each well.]] | ||
:* Colony PCR for GFP fusion brick and new pRL1383a to verify inserts | :* Colony PCR for GFP fusion brick and new pRL1383a to verify inserts | ||
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::* BBa_J04430 | ::* BBa_J04430 | ||
::* BBa_J33207 | ::* BBa_J33207 | ||
| - | ::* | + | ::* BBa_R001 |
:* Ran all PCR products on a 4% gel to check | :* Ran all PCR products on a 4% gel to check | ||
::* Gel ran funny -- loading dye ran 2x as fast as the DNA itself | ::* Gel ran funny -- loading dye ran 2x as fast as the DNA itself | ||
| + | :::* 119V too much to handle? Wavy bands | ||
::* Poor ladder resolution on gel | ::* Poor ladder resolution on gel | ||
::* No bands seen for GFP fusion, pRL1383a, R0010 | ::* No bands seen for GFP fusion, pRL1383a, R0010 | ||
| - | ::* Incorrect bands for B0015 and J33207 | + | ::* Incorrect bands for B0015 (multiple bands, what are the two bands at the top?) and J33207 (size too small, stuff still in wells) |
::* Will redo plasmid preps for R0010, B0015, J33207 | ::* Will redo plasmid preps for R0010, B0015, J33207 | ||
::* Will redo PCR reaction tomorrow for pRL1383a, GFP fusion, B0015, J33207, R0010 | ::* Will redo PCR reaction tomorrow for pRL1383a, GFP fusion, B0015, J33207, R0010 | ||
Revision as of 04:47, 18 July 2008
| Projects | Events | Resources | ||
|---|---|---|---|---|
| Sponsors | Experiments | Milestones | Protocols | |
| Notebook (t) | Meetings (t) |
Things we did today
Wetlab work
PCR
- Grace
- Colony PCR for GFP fusion brick and new pRL1383a to verify inserts
- Sequencing (25 mu;l reactions)
- nir promoter
- slr2016
- pilA
- BBa_B0015
- BBa_B0024
- BBa_B0030
- BBa_C0012
- BBa_E0040
- BBa_J04430
- BBa_J33207
- BBa_R001
- Ran all PCR products on a 4% gel to check
- Gel ran funny -- loading dye ran 2x as fast as the DNA itself
- 119V too much to handle? Wavy bands
- Poor ladder resolution on gel
- No bands seen for GFP fusion, pRL1383a, R0010
- Incorrect bands for B0015 (multiple bands, what are the two bands at the top?) and J33207 (size too small, stuff still in wells)
- Will redo plasmid preps for R0010, B0015, J33207
- Will redo PCR reaction tomorrow for pRL1383a, GFP fusion, B0015, J33207, R0010
Plasmid Prep
- Grace
- Inoculated 7 ml LB + amp100 with single colony of B0015, R0010, J04430 for redo of plasmid prep tomorrow
Restreak to Purify
- Grace
- Restreaked GFP fusion and pRL1383a (MCS replaced) transformants to purify
pSB3K3 plasmid prep
- Margaret
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 
]
