From 2014.igem.org
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| - | <ul><li>PCR'ed out LacI and TetR genes from pAA001 plasmid due to sequence similarity problems</li> | + | To figure out what went wrong during assembly of the combinatorial promoter constructs used in last week's TXTL workshop, we attempted to take out the LacI and TetR genes present in plasmids pAA001 and pAA002. |
| - | <li>Inserted new pAA001 plasmids into competent cells and plated</li> | + | <ul><li>PCR extraction of linear fragments of pAA001 and pAA002 minus the lacI and tetR genes.</li> |
| | + | <li>Recircularization of the truncated "backbone"+"combinatorial promoter construct" round-the-horn PCR</li> |
| | + | <li>Transformation of recircularized plasmids into JM109 competent cells</li> |
| | + | <li>Cells plated onto carbenicillin plates and incubated at 37°C overnight</li> |
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| | + | In other news, we continued to work on the wiki and make it worse and worse...... |
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| - | <ul><li>Picked 12 colonies from yesterday's transformed cells and performed colony PCR</li> | + | <ul><li></li> |
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Revision as of 22:40, 3 July 2014
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Notebook |
Overview
Week 1
Week 2
Week 3
Week 4
Week 5
Week 6
Week 7
Week 8
Week 9
Week 10
Week 11
Week 12
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Week Three |
Monday, 6/30/14
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- Discussion of fallout from Friday's meeting
- More paper-reading to determine what's wrong with our new quorum-sensing system candidates
- Designed geneblocks for our new systems and subgroup projects
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Tuesday, 7/1/14
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- Lab cleanup. Reorganized -20 freezer samples.
- World Cup watching. Tim Howard's a boss
- Work on the team wiki while we wait for our next batch of geneblocks to ship
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Wednesday, 7/2/14
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To figure out what went wrong during assembly of the combinatorial promoter constructs used in last week's TXTL workshop, we attempted to take out the LacI and TetR genes present in plasmids pAA001 and pAA002.
- PCR extraction of linear fragments of pAA001 and pAA002 minus the lacI and tetR genes.
- Recircularization of the truncated "backbone"+"combinatorial promoter construct" round-the-horn PCR
- Transformation of recircularized plasmids into JM109 competent cells
- Cells plated onto carbenicillin plates and incubated at 37°C overnight
In other news, we continued to work on the wiki and make it worse and worse......
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Thursday, 7/3/14
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Friday, 7/4/14
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It's the 4th of July!!!
'Murica...or something like that, I guess...
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"
