|
|
| Line 360: |
Line 360: |
| | description: '<b>Members:</b> Rob, Andrew <br> <b>What we did: </b>Final chloride assay showed convincing degradation of chloroacetate and DCA using the parts we submitted to the iGEM HQ, also showed that our inducible-promoter systems failed to assemble correctly, instead containing just the constitutive promoter (Ptet). Andrew found further evidence for this by PCR screening.<br><br>Cleaned and packed up a lot of stuff in the lab. ' | | description: '<b>Members:</b> Rob, Andrew <br> <b>What we did: </b>Final chloride assay showed convincing degradation of chloroacetate and DCA using the parts we submitted to the iGEM HQ, also showed that our inducible-promoter systems failed to assemble correctly, instead containing just the constitutive promoter (Ptet). Andrew found further evidence for this by PCR screening.<br><br>Cleaned and packed up a lot of stuff in the lab. ' |
| | }, | | }, |
| | + | |
| | { | | { |
| - | title: 'Wikifreeze', | + | title: 'Ignore this, I\'m testing :)', |
| - | start: new Date(2013, 8, 27),
| + | start: new Date(y, m, 28), |
| - | description: '<b>Members:</b> Everyone <br> <b>What we did: </b> Ohh no!! Wikifreeze! '
| + | end: new Date(y, m, 29), |
| - | },
| + | url: 'http://google.com/' |
| - | {
| + | |
| - | title: 'Hong Kong Jamboree Preparation',
| + | |
| - | start: new Date(2013, 8, 30),
| + | |
| - | description: '<b>Members:</b> Everyone <br> <b>What we did: </b> Preparing our poster and presentation for HK. '
| + | |
| - | },
| + | |
| - | {
| + | |
| - | title: 'Hong Kong Jamboree Preparation',
| + | |
| - | start: new Date(2013, 9, 1), | + | |
| - | description: '<b>Members:</b> Everyone <br> <b>What we did: </b> Preparing our poster and presentation for HK. ' | + | |
| - | },
| + | |
| - | {
| + | |
| - | title: 'Hong Kong Jamboree Preparation',
| + | |
| - | start: new Date(2013, 9, 2),
| + | |
| - | description: '<b>Members:</b> Everyone <br> <b>What we did: </b> Preparing our poster and presentation for HK. ' | + | |
| - | },
| + | |
| - | {
| + | |
| - | title: 'Hong Kong Jamboree',
| + | |
| - | start: new Date(2013, 9, 4),
| + | |
| - | description: '<b>Members:</b> Everyone <br> <b>What we did: </b> Practice presentation until the early hours. '
| + | |
| - | },
| + | |
| - | {
| + | |
| - | title: 'Hong Kong Jamboree',
| + | |
| - | start: new Date(2013, 9, 5),
| + | |
| - | description: '<b>Members:</b> Everyone <br> <b>What we did: </b> Presentations all day! '
| + | |
| - | },
| + | |
| - | {
| + | |
| - | title: 'Hong Kong Jamboree',
| + | |
| - | start: new Date(2013, 9, 6),
| + | |
| - | description: '<b>Members:</b> Everyone <br> <b>What we did: </b> Won the Best Human Practices and Best Experimental Measurement Approach! '
| + | |
| - | },
| + | |
| - | {
| + | |
| - | title: 'Planning post-HK labwork',
| + | |
| - | start: new Date(2013, 9, 9),
| + | |
| - | description: '<b>Members:</b> Rob, Andrew <br> <b>What we did: </b> Designing gBlocks and primers in Ho Chi Minh airport for labwork before MIT.'
| + | |
| - | },
| + | |
| - | {
| + | |
| - | title: 'Ordering new gBlocks',
| + | |
| - | start: new Date(2013, 9, 10),
| + | |
| - | description: '<b>Members:</b> Rob, Andrew <br> <b>What we did: </b> Convincing our supervisor to purchase the new gBlocks and primers we need. '
| + | |
| - | },
| + | |
| - | {
| + | |
| - | title: 'Finding dhlB',
| + | |
| - | start: new Date(2013, 9, 11),
| + | |
| - | description: '<b>Members:</b> Hugh, Shuravi <br> <b>What we did: </b> Transformation of more clones with pSB1C3-dhlB ligation mixture. PCR Screen for correct orientation of dhlB from non-directional ligation. '
| + | |
| - | },
| + | |
| - | {
| + | |
| - | title: 'Isolating dhlB',
| + | |
| - | start: new Date(2013, 9, 12),
| + | |
| - | description: '<b>Members:</b> Andrew, Rob <br> <b>What we did: </b> Plasmid prep of promising clones containing pSB1C3-dhlB.'
| + | |
| - | },
| + | |
| - | {
| + | |
| - | title: 'Confirming dhlB',
| + | |
| - | start: new Date(2013, 9, 13),
| + | |
| - | description: '<b>Members:</b> Andrew <br> <b>What we did: </b> Digest confirmation of length and concentration of pSB1C3-dhlB.'
| + | |
| - | },
| + | |
| - | {
| + | |
| - | title: 'Cloning dhlB',
| + | |
| - | start: new Date(2013, 9, 14),
| + | |
| - | description: '<b>Members:</b> Rob <br> <b>What we did: </b> Cloning of the promoter Ptet (PCR product) into pSB1C3-dhlB.'
| + | |
| - | },
| + | |
| - | {
| + | |
| - | title: 'Confirming expression of dhlB',
| + | |
| - | start: new Date(2013, 9, 15),
| + | |
| - | description: '<b>Members:</b> Rob, Desmond <br> <b>What we did: </b> Plating pSB1C3-Ptet-dhlB clones on pH-chloroacetate plates to find clones degrading chloroacetate. Set-up an overnight incubation of resting cells to confirm chloroacetate degradation.'
| + | |
| - | }, {
| + | |
| - | title: 'Further confirmation',
| + | |
| - | start: new Date(2013, 9, 16),
| + | |
| - | description: '<b>Members:</b> James <br> <b>What we did: </b> Chloride assay to confirm chloroacetate degradation. Woo! We’ve got dhlB isolated and it works!'
| + | |
| - | }, {
| + | |
| - | title: 'gBlocks arrived!',
| + | |
| - | start: new Date(2013, 9, 17),
| + | |
| - | description: '<b>Members:</b> Rob, Andrew <br> <b>What we did: </b> gBlocks for p450, aldA and tetR arrived. Gibson Assembly and transformation.
| + | |
| - | '
| + | |
| - | }, {
| + | |
| - | title: 'Gibson transformation #1',
| + | |
| - | start: new Date(2013, 9, 18),
| + | |
| - | description: '<b>Members:</b> Andrew <br> <b>What we did: </b> Checked plates after Gibson Assembly - no colonies on tests and colonies on negative controls. Suspect bad plates. Remade plates and re-transformed Gibson reaction product. '
| + | |
| - | },
| + | |
| - | {
| + | |
| - | title: 'PCR screening',
| + | |
| - | start: new Date(2013, 9, 19),
| + | |
| - | description: '<b>Members:</b> Rob <br> <b>What we did: </b> Checked plates again - better! Set-up a huge PCR screen for all Gibson constructs - aldA, p450 and tetR. Retransformed p450 Gibson reaction product to generate more clones for screening. '
| + | |
| - | },
| + | |
| - | {
| + | |
| - | title: 'Plasmid prep',
| + | |
| - | start: new Date(2013, 9, 20),
| + | |
| - | description: '<b>Members:</b> Rob, Andrew, Shuravi <br> <b>What we did: </b> More screening. Plasmid prep of promising clones. '
| + | |
| - | },
| + | |
| - | {
| + | |
| - | title: 'Cloning',
| + | |
| - | start: new Date(2013, 9, 21),
| + | |
| - | description: '<b>Members:</b> Rob, Andrew, Hugh <br> <b>What we did: </b> Digest confirmation of plasmid with a RE specific to each gBlock. Confirmed length and concentration of all new parts. Digestion, gel purification, ligation and transformation of new parts so that they can be expressed. '
| + | |
| - | },
| + | |
| - | {
| + | |
| - | title: 'Waiting for growth',
| + | |
| - | start: new Date(2013, 9, 22),
| + | |
| - | description: '<b>Members:</b> Andrew <br> <b>What we did: </b> Transformation plates hadn’t grown sufficiently for screening. '
| + | |
| - | },
| + | |
| - | {
| + | |
| - | title: 'Screening clones',
| + | |
| - | start: new Date(2013, 9, 23),
| + | |
| - | description: '<b>Members:</b> Rob, Hugh, Andrew <br> <b>What we did: </b> Patching and lysing cells from transformation plates for PCR screen and pH-chloroacetate screens. No efficient cloning for whole pathway, only for individual parts with tetR-inducible promoter system. Set-up chloride assay for degradation of DCA by clones containing tetR-p450 and tetR-aldA-dhlB-dhlA.'
| + | |
| - | },
| + | |
| - | {
| + | |
| - | title: 'Chloride assay',
| + | |
| - | start: new Date(2013, 9, 24),
| + | |
| - | description: '<b>Members:</b> Rob, Andrew <br> <b>What we did: </b> Performed chloride assay. Appears there is reasonable degradation of DCA by p450.
| + | |
| - | '
| + | |
| - | },
| + | |
| - | {
| + | |
| - | title: 'Acetaldehyde growth inhibition',
| + | |
| - | start: new Date(2013, 9, 25),
| + | |
| - | description: '<b>Members:</b> Rob, Andrew <br> <b>What we did: </b> Set-up an experiment to confirm aldA expression through growth inhibition of control E. coli in LB-acetaldehyde.'
| + | |
| - | },
| + | |
| - | {
| + | |
| - | title: 'Characterising new parts',
| + | |
| - | start: new Date(2013, 9, 26),
| + | |
| - | description: '<b>Members:</b> Rob, Andrew <br> <b>What we did: </b> Another LB-acetaldehyde growth experiment in triplicate with a promising tetR-aldA clone. Incubated tetR-p450 clones with DCA in triplicate to confirm result from the 24th October, after 6 hours incubation, GC was not promising. Set-up an ethene-epoxide assay to confirm p450 expression. '
| + | |
| | } | | } |
| | ] | | ] |
"
