Template:Team:SydneyUni Australia/Calendar/Events List

From 2013.igem.org

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start: new Date(2013, 8, 15),
start: new Date(2013, 8, 15),
description: '<b>Members:</b> Rob, Andrew, James <br> <b>What we did: </b>Following on from yesterdays digested plasmid prep a gel was run to at different concentrations.<br><br>We found, from the gel of our digest, that the plasmids were not concentrated enough.<br><br>Another plasmid prep, using some different clones was done. This time however, we incubated cells overnight to increase plasmid yield.<br><br>We digested and ran a gel of our plasmid for confirmation. <br><br><div style=\'color:red;font-weight:bold;\'>The gel turned out perfect!</div>'
description: '<b>Members:</b> Rob, Andrew, James <br> <b>What we did: </b>Following on from yesterdays digested plasmid prep a gel was run to at different concentrations.<br><br>We found, from the gel of our digest, that the plasmids were not concentrated enough.<br><br>Another plasmid prep, using some different clones was done. This time however, we incubated cells overnight to increase plasmid yield.<br><br>We digested and ran a gel of our plasmid for confirmation. <br><br><div style=\'color:red;font-weight:bold;\'>The gel turned out perfect!</div>'
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},
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{
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title: 'Sending off Parts',
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start: new Date(2013, 8, 16),
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description: '<b>Members:</b> Viv, James, Andrew, Rob <br> <b>What we did: </b>We sent off our parts for dhlA (BBa_K1115004), dhlB (BBa_K1115005) and dhlB-dhlA (BBa_K1115006), based on the results from PCR screening of cells and digests of plasmid preps.<br><br>However, we did a PCR screen using the plasmids as templates and discovered one of our parts is a reverse insert! This is dhlB (BBa_K1115005).'
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},
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{
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title: 'Sending off Parts, Characterisation',
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start: new Date(2013, 8, 17),
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description: '<b>Members:</b> Andrew, Rob <br> <b>What we did: </b>Re-submitted parts we’re pretty sure are correct. Started setting up cells and reagents for construction of a constitutive and inducible promoter system to characterise our parts in pSB1C#3 before the wiki-freeze.<br><br>PCR of parts directly from Distribution Kit for subsequent use in directional cloning.'
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},
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{
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title: 'Transformation and Promoter Hunting',
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start: new Date(2013, 8, 18),
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description: '<b>Members:</b> Rob, Shuravi, Viv, Andrew <br> <b>What we did: </b>Construction of Pcat-dhlB-dhlA in pSB1C3. Rob wrote out all the protocol and ran a digest, Viv ligated and transformed the construct, Shuravi spread-plated colonies.<br><br>Andrew went through a lot of troubleshooting trying to PCR parts or plasmid prep parts from the Distribution Kit for a lacI inducible promoter.'
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},
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{
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title: 'Screening Plates',
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start: new Date(2013, 8, 19),
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description: '<b>Members:</b> Rob, Andrew <br> <b>What we did: </b>Figured out we could make screening plates to help us find transformants that are expressing our construct. We added chloroacetate and phenol red to LB-agar before plating and looked for colour change as the bacteria degraded chloroacetate and released Cl- ions.'
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},
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{
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title: 'Promoter and Plate Construction',
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start: new Date(2013, 8, 20),
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description: '<b>Members:</b> Rob, Andrew <br> <b>What we did: </b>Construction of inducible promoter system in pSB1C3. Ran out of luck with lacI and turned to arabinose and tetracycline systems. PCR of parts from the Distribution Kit and verification of length and purity on gels.<br><br>Started playing around with the ingredients of chloroacetate-phenol red screening plates. We did a quick titration to find an optimal pH close to the colour-change from phenol red. '
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},
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{
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title: 'Promoter Work, Selection Plates',
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start: new Date(2013, 8, 21),
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end: new Date(2013, 8, 22),
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description: '<b>Members:</b> Rob, Andrew, James, Viv <br> <b>What we did: </b>Sequential digestion and ligation of PCR products for an inducible promoter (Ptet and TetR, Pbad and AraC, parts from Distribution Kit).<br><br>Found that LB-agar-chloramphenicol plates with 10mM chloroacetate, 18mg/L phenol red, pH 6.8, worked best and allowed us to pick a few clones constitutively expressing dhlB for further characterisation by chloride assay.'
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},
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{
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title: 'Chloride Assay, Inducibility Screen',
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start: new Date(2013, 8, 23),
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description: '<b>Members:</b> Rob, Andrew, Viv <br> <b>What we did: </b>Set-up chloride assay for a few promising clones constitutively expressing dhlB.<br><br>Set-up plates using the same screening system to find clones with inducible expression of dhlB.'
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},
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{
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title: 'Chloride Assay',
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start: new Date(2013, 8, 24),
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description: '<b>Members:</b> Rob, Andrew <br> <b>What we did: </b>Chloride assay showed degradation of chloroacetate and DCA using the parts we submitted to the iGEM HQ.'
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},
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{
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title: 'Triplicate Chloride Assay',
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start: new Date(2013, 8, 25),
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description: '<b>Members:</b> Rob, Andrew, James <br> <b>What we did: </b>Set-up another chloride assay in triplicate for neat characterisation of our parts, in parallel with promising clones with inducible-promoter systems.<br><br>James ran an SDS-page gel for further evidence that our parts are being expressed.'
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},
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{
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title: 'Chloride Assay, Promoter Characterisation',
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start: new Date(2013, 8, 26),
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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. '
},
},

Revision as of 06:34, 27 September 2013

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