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 Eleven |
Monday, 8/25/14
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lamBCDA & fsrABC Reception Systems
agrBCDA Reception System and Combinatorial Promoters
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Tuesday, 8/26/14
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lamBCDA & fsrABC Reception Systems
agrBCDA Reception System and Combinatorial Promoters
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Wednesday, 8/27/14
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lamBCDA & fsrABC Reception Systems
agrBCDA Reception System and Combinatorial Promoters
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Thursday, 8/28/14
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lamBCDA & fsrABC Reception Systems
agrBCDA Reception System and Combinatorial Promoters
- Re-ran a Gibson assembly of pAA009
- Transformed Gibson products into JM109 cells
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Friday, 8/29/14
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lamBCDA & fsrABC Reception Systems
agrBCDA Reception System and Combinatorial Promoters
- Picked 10 Gibson colonies from yesterday
- Ran colony PCR on re-suspended colonies, inoculated liquid cultures of picked colonies
- Ran gel of colony PCR products, but no good bands were obtained
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Saturday, 8/30/14
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Export Systems
Redo of cloning of agr system:
- Ran PCR reactions to construct appropriate backbone for pTG002 & modified gblock insert for pTG002.
- DpnI digested PCR product for pTG002 backbone.
- PCR purified DpnI digested pTG002 backbone and pTG002 modified gblock insert PCR products.
- Ran purified products on a gel.
- Ran Gibson assembly reaction between pTG002-backbone & pTG002-insert.
- Transformed Gibson reaction mix into JM109. Cells were then plated & incubated overnight.
De-FLAGging pTG005 for LC/MS analysis:
- Ran PCR reaction to exclude 3xFLAG from pTG005.
- Ran DpnI digest and then PCR purification on PCR product.
- Ran PCR product on a gel to verify creation of the appropriate DNA fragment.
- Ran a round-the-horn ligation reaction on the purified PCR product.
- Transformed the reaction mix into JM109. Cells were then plated & incubated overnight.
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