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
Week 13
Week 14
Week 15
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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, B83 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.
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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