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 Six |
Monday, 7/21/14
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Export Systems
3 of our 5 constructs were not properly cloned, so we are going to redo them:
- PCRed out backbones for pTG002, pTG003, and pTG006 with overhangs
- PCR product was then DpnI digested and PCR-purified. A gel was run with processed and unprocessed product to verify backbone creation
- Gibson assembly of pTG002, pTG003, and pTG006 (pTG006 had 2 reactions: 1 with the backbone created last week, since it had been verified to work for cloning pTG005, and another with backbone created today)
- Assemblies were transformed into JM109, plated on carbenicillin plates, and incubated overnight
lamBCDA & fsrABC Reception Systems
- 4 cultures of JM109 were transformed, plated on Kan + Cm plates, and incubated overnight at 37°C:
- pMB001 and pMB003
- pMB002 and pMB003
- pMB004 and pMB006
- pMB005 and pMB006
agrBCDA Reception System and Combinatorial Promoters
For the agrBCDA System, we troubleshot our problems creating the pAA009/10 vector backbone:
- Ran a gel of the PCR product created on Friday (pAA009/10 backbone). Results were negative.
- Tried PCR again with lower annealing temperatures [(what temps?)]. Results were negative again.
For the combinatorial promoters:
- Set up overnight culture of jwAA001 strain to do more experiments tomorrow
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Tuesday, 7/22/14
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Wednesday, 7/23/14
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Thursday, 7/24/14
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Friday, 7/25/14
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