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Virginia Commonwealth/26 June 2009
From 2009.igem.org
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*1 Pick colonies and grow overnight. | *1 Pick colonies and grow overnight. | ||
*2 Confirm the growing colonies using GFP and RFP reporter. | *2 Confirm the growing colonies using GFP and RFP reporter. | ||
| - | *3 Order DNA | + | *3 Order DNA and IPTG |
| + | *4 Additional promoters were designed. These possessed an up element that is proven to bind with the alpha subunit of RNAP by multiple sources. | ||
====Wetlab==== | ====Wetlab==== | ||
*1 IPTG stock was prepared. Since only 0.746g of IPTG stock was available, only 2.64 mL was prepared. - | *1 IPTG stock was prepared. Since only 0.746g of IPTG stock was available, only 2.64 mL was prepared. - | ||
*2 IPTG stock (5.0 microliters) was added to 5 mL of bacteria that was grown overnight with pSB3k3 backbone (to induce the expression of RFP). After several hours no fluorescence was expressed. 3 mL LB+KAN was added to each culture so that the cells could re-enter log phase growth. 3 uL IPTG was also added to each culture to maintain concentration. | *2 IPTG stock (5.0 microliters) was added to 5 mL of bacteria that was grown overnight with pSB3k3 backbone (to induce the expression of RFP). After several hours no fluorescence was expressed. 3 mL LB+KAN was added to each culture so that the cells could re-enter log phase growth. 3 uL IPTG was also added to each culture to maintain concentration. | ||
Revision as of 18:07, 26 June 2009
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Contents |
Friday 26 June 2009
Results
- 1 Plate growing pSB3K3 plasmid grew well. There was no fluorescence expressed.
- 2 Plate growing the device plasmid pSB3K3, BBa_J23102, and E0240 grew well. There was no fluorescence expressed.
Tasks
- 1 Pick colonies and grow overnight.
- 2 Confirm the growing colonies using GFP and RFP reporter.
- 3 Order DNA and IPTG
- 4 Additional promoters were designed. These possessed an up element that is proven to bind with the alpha subunit of RNAP by multiple sources.
Wetlab
- 1 IPTG stock was prepared. Since only 0.746g of IPTG stock was available, only 2.64 mL was prepared. -
- 2 IPTG stock (5.0 microliters) was added to 5 mL of bacteria that was grown overnight with pSB3k3 backbone (to induce the expression of RFP). After several hours no fluorescence was expressed. 3 mL LB+KAN was added to each culture so that the cells could re-enter log phase growth. 3 uL IPTG was also added to each culture to maintain concentration.
