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| Line 42: |
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| | start: new Date(2013, 4, 24), | | start: new Date(2013, 4, 24), |
| | description: '<b>Members:</b> Coleman, Viv, Rob <br> <b>What we did:</b> Finished the ToMO plasmid prep. Digested pBBR1-MCS2. Used the nanodrop. Did a digest + gel of our ToMO, undigested and Xbal1-digested pBBR along with a marker' | | description: '<b>Members:</b> Coleman, Viv, Rob <br> <b>What we did:</b> Finished the ToMO plasmid prep. Digested pBBR1-MCS2. Used the nanodrop. Did a digest + gel of our ToMO, undigested and Xbal1-digested pBBR along with a marker' |
| - | },
| |
| - | // -----------------Week 2 ----------------
| |
| - | {
| |
| - | title: 'Transformation and Selection',
| |
| - | start: new Date(2013, 4, 29),
| |
| - | description: '<b>Members:</b> Coleman, Cyril, Shuravi, Rob <br> <b>What we did:</b> We transformed E. coli Epi300 with two plasmids, pBBR and pBS(ToMO). As both have Km resistance, only those cells that were successfully transformed with the plasmid would grown when plated onto LB+Km media. They were incubated overnight'
| |
| - | },
| |
| - | {
| |
| - | title: 'Growing up cells',
| |
| - | start: new Date(2013, 4, 30),
| |
| - | description: '<b>Members:</b> Coleman, Andrew, Desmond <br> <b>What we did:</b> The two E. Coli treatments were growing at different rates, E. Coli (pBS-ToMO) wasn’t growing well at all. We transferred E. Coli (pBBR) to broth and later washed and transferred to two phosphate buffer treatments, one with DCA, one without. We transferred E. Coli (pBS-ToMO) to broth and incubated it overnight. DCA concentration was 1mM. '
| |
| - | },
| |
| - | {
| |
| - | title: 'Chloride Assay',
| |
| - | start: new Date(2013, 4, 31),
| |
| - | description: '<b>Members:</b> Colman, Rob, Hugh<br> <b>What we did:</b> After 24 hours in DCA, we put E. Coli (pBBR) in the cool room for a Cl assay next week. We washed and transferred E. Coli (pBS-ToMO) from broth to two phosphate buffer treatments, one with DCA, one without. We also ran a quick PCR testing our new ToMO primers on pBS-ToMO, but we didn’t digest pBS-ToMO before PCR. !'
| |
| - | },
| |
| - | // -----------------Week 3 ----------------
| |
| - | {
| |
| - | title: 'PCR analysis and GC analysis of DCA metabolism',
| |
| - | start: new Date(2013, 5, 3),
| |
| - | description: '<b>Members:</b> Colman, Hugh, Rob <br> <b>What we did:</b> We ran a gel of last Friday’s PCR. The PCR was of pBS(ToMO) extracted from Wood’s E. Coli TG1, with primers iGEM 1 & 2. The banding on the gel was obscure, potentially due to too much template and could not be used. <br><br> We used the Gas Chromatographer (GC) to look at our two DCA treatments of E. Coli (transformed with either pBBR or pBS(ToMO)). Both had pretty much the same reading but it is uncertain yet whether this is because pBS(ToMO) was lost or altered during the extended incubation from Week 2, or whether ToMO was unable to degrade DCA. We also set up standardised solutions of NaCl in KP buffer for a future Cl assay.'
| |
| - | },
| |
| - | // -----------------Week 4 ----------------
| |
| - | {
| |
| - | title: 'Redo of ToMO Trial',
| |
| - | start: new Date(2013, 5, 22),
| |
| - | end: new Date(2013, 5, 27),
| |
| - | description: '<b>Members:</b> Coleman, Rob <br> <b>What we did:</b> We repeated the ToMO trial however made some changes to the protocol. '
| |
| | }, | | }, |
| - |
| |
| | | | |
| | { | | { |
"
