Sandbox: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
No edit summary |
No edit summary |
||
Line 32: | Line 32: | ||
|'''Preliminary wet work ''' || || || | |'''Preliminary wet work ''' || || || | ||
|- | |- | ||
| ||Extract promoter, RBS and terminator BioBricks from registry | | ||Extract promoter, RBS and terminator BioBricks from registry || || | ||
:*Refine protocol for paper-bound DNA extraction | | ||:*Refine protocol for paper-bound DNA extraction || || | ||
:*Use PCR and transformations to confirm presence of DNA | | ||:*Use PCR and transformations to confirm presence of DNA || || | ||
|| || | |||
|- | |- | ||
|'''Internal K+ build-up''' || || || | |'''Internal K+ build-up''' || || || |
Revision as of 08:32, 4 August 2008
<html>
<td class="tdboxes"><a href="http://openwetware.org/wiki/iGEM:Cambridge/2008/Notebook/Voltage"><img border="0" src="http://openwetware.org/images/0/00/Vol_logo.JPG" width="256" height="201"></a></td>
<td class="tdboxes"><a href="http://openwetware.org/wiki/iGEM:Cambridge/2008/Notebook/Voltage"><img border="0" src="http://openwetware.org/images/d/d1/Mag_logo.JPG" width="210" height="201"></a></td>
<td class="tdboxes"><a href="http://openwetware.org/wiki/iGEM:Cambridge/2008/Notebook/Voltage"><img border="0" src="http://openwetware.org/images/f/fe/Tur_logo.JPG" width="201" height="201"></a></td>
</html>
Personnel | Progress | ||||||||||
Research | |||||||||||
Potassium intake | |||||||||||
Preventing K+ efflux | |||||||||||
Bacterial tolerance for high K+ and turgor
|
|||||||||||
Ligand gated channels | |||||||||||
Media | |||||||||||
Preliminary wet work | |||||||||||
Extract promoter, RBS and terminator BioBricks from registry | :*Refine protocol for paper-bound DNA extraction | :*Use PCR and transformations to confirm presence of DNA | |||||||||
Internal K+ build-up | |||||||||||
PCR Kdp K+ pump gene from E.coli MG1655
|
|||||||||||
Put Kdp gene under control of stationary phase promoter (osmY, used by MIT 2006 team)
|
|||||||||||
Transform into wildtype and mutant E.coli strains | |||||||||||
Test
|
|||||||||||
Chassis | |||||||||||
Order from Yale
| |||||||||||
Test
| |||||||||||
Controlled K+ efflux | |||||||||||
Design sequence based on GluR0 glutamate-gated K+ channel from Synechocystis PCC 6803 | |||||||||||
Send to DNA 2.0 for synthesis | |||||||||||
Backup
| |||||||||||
Ligate gene into BioBrick plasmid | |||||||||||
Transform into chosen chassis | |||||||||||
Test
|
|||||||||||
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 |