Team:Cambridge/Notebook/Week4

From 2010.igem.org

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==Tuesday==
==Tuesday==
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* Wet Work
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We made cells from 4 different hns mutants: W3110 hns93-1, BW25113 Δhns::kan, W3110 hns-205::Tn10, GM230 hns-205::Tn10.  This was prompted by a paper suggesting increased bioluminescence in strains with reduced hns. These cells were stored at -80°C for later use
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** Made competent cells of 4 different hns mutants: W3110 hns93-1, BW25113 Δhns::kan, W3110 hns-205::Tn10, GM230 hns-205::Tn10
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** Now stored in the -80°C freezer for later use
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* Dry Work
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We continued to discuss synthesis requirements
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** Organising sequences for synthesis
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{{Team:Cambridge/Templates/Day|Day=Wednesday}}
{{Team:Cambridge/Templates/Day|Day=Wednesday}}
{{:Team:Cambridge/Templates/rightpic|src=Cambridge-Gel.jpg}}
{{:Team:Cambridge/Templates/rightpic|src=Cambridge-Gel.jpg}}
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*Hannah and Theo tested previous day's preparations for competence
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Hannah and Theo tested the previous day's preparations for competence.  Unfortunately they plated out only 20ul, not enough to get sufficient colonies for an accurate perception of competence. Meanwhile, Peter and Paul transformed E.coli containing phK555 with phK724 and placed the cells in a 30C incubator. (V. fischeri lux proteins denature above this temperature)
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*Peter and Paul transformed phK555 with phK724, placed cells in 30C incubator (above rotating incubator)
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*Bill, Emily, Anja performed miniprep and restriction digest then ligation.
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**Decided that we may have used wrong technique, but DNA samples in fridge
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Bill, Emily, Anja tried to add a prokaryotic RBS and promoter to the firefly luciferase already in the registry.  They performed miniprep and restriction digest then ligation, but used incorrect restriction enzymes, such that we could not exclude religated plasmid backbones.
{{Team:Cambridge/Templates/Day|Day=Thursday}}
{{Team:Cambridge/Templates/Day|Day=Thursday}}
{{:Team:Cambridge/Templates/rightpic|src=Cambridge-Glow.jpg}}
{{:Team:Cambridge/Templates/rightpic|src=Cambridge-Glow.jpg}}

Revision as of 17:08, 14 August 2010