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 Two |
Monday, 6/23/14
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- Discussion of fallout from Friday's meeting, paper reading
- Colony PCR and Minipreps of liquid cultures of colonies picked over the weekend
- TXTL workshop starts today
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Tuesday, 6/24/14
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- Discussed alternatives to current ComQXPA system, including 2-cell system (not all in 1 cell)
- Gibson assembly of another combinatorial promoter using pKS001 backbone, lacI geneblock, ____ promoter geneblock, and sfGFP geneblock.
- Transformation of Gibson constructs into JM109 cells
- Linear combinatorial constructs created via PCR of pAA001 (minipreps) to be tested in TXTL
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Wednesday, 6/25/14
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- Linear constructs and plasmid TXTL reactions of A90 promoter (pAA001) were set up and run
- Gibson assembly of plasmid pAA002 containing B83 promoter
- PCR of Gibson assemblies of pAA002 to create linear constructs to test in TXTL
- TXTL reactions of B83 promoter (linear frag. from pAA002) were set up and run
- Transformation of pAA002 Gibson assemblies into JM109
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Thursday, 6/26/14
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- Analysis of yesterday's TXTL reactions (run overnight)
- Colony PCR and miniprep of bacterial colonies transformed with pAA002. Minipreps shipped for sequencing.
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TXTL reaction data inconclusive. Most fluorescence signals are below negative control, and linear data frequently contradicts plasmid data with A90 promoter. No signal observed for B83 promoter. |
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Friday, 6/27/14
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- Continued to investigate different signalling pathways
- Potential alternative pathways to investigate:
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