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Team:Caltech/week11
From 2014.igem.org
Revision as of 00:53, 15 September 2014 by
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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
Week Eleven
Monday, 8/25/14
lamBCDA & fsrABC Reception Systems
agrBCDA Reception System and Combinatorial Promoters
Tuesday, 8/26/14
lamBCDA & fsrABC Reception Systems
agrBCDA Reception System and Combinatorial Promoters
Wednesday, 8/27/14
lamBCDA & fsrABC Reception Systems
agrBCDA Reception System and Combinatorial Promoters
Thursday, 8/28/14
lamBCDA & fsrABC Reception Systems
agrBCDA Reception System and Combinatorial Promoters
Re-ran a Gibson assembly of pAA009
Transformed Gibson products into JM109 cells
Friday, 8/29/14
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
Saturday, 8/30/14
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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