Sandbox: Difference between revisions
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|'''Preliminary wet work ''' || || || | |'''Preliminary wet work ''' || || || | ||
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| ||Extract promoter, RBS and terminator BioBricks from registry | | ||Extract promoter, RBS and terminator BioBricks from registry | ||
:*Refine protocol for paper-bound DNA extraction | |||
:*Use PCR and transformations to confirm presence of DNA | |||
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|'''Internal K+ build-up''' || || || | |'''Internal K+ build-up''' || || || |
Revision as of 08:37, 4 August 2008
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Personnel | Progress | ||
Research | |||
Potassium intake | |||
Preventing K+ efflux | |||
Bacterial tolerance for high K+ and turgor
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Ligand gated channels | |||
Media | |||
Preliminary wet work | |||
Extract promoter, RBS and terminator BioBricks from registry
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Internal K+ build-up | |||
PCR Kdp K+ pump gene from E.coli MG1655
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Put Kdp gene under control of stationary phase promoter (osmY, used by MIT 2006 team)
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Transform into wildtype and mutant E.coli strains | |||
Test
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Chassis | |||
Order from Yale
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Test
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Controlled K+ efflux | |||
Design sequence based on GluR0 glutamate-gated K+ channel from Synechocystis PCC 6803 | |||
Send to DNA 2.0 for synthesis | |||
Backup
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Ligate gene into BioBrick plasmid | |||
Transform into chosen chassis | |||
Test
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Measuring voltage | |||
Quantify output using oxygen electrode or glass capillary microelectrode | |||
Medium optimisation | |||
Vary K+ concentrations, using KCl | |||
Vary nutrient levels | |||
Output optimisation | |||
Vary strength of promoters/RBS |