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Team:KULeuven/15 July 2008
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== Lab Work == | == Lab Work == | ||
Revision as of 07:12, 16 July 2008
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Contents |
Lab Work
Wetlab
Today was our first day in the lab. We prepared LB-medium, Ampicilline,... Everything is now ready for the serious labwork: from tips to cleaning our benches. Today was also a historic date, because our big scheme was born! (see Parts-page) We did a first attempt to punch out a part of the registry. Then we transformed competent cells (DH5alpha) with the part. We started with the input device: M30109. Stefanie was very excited to pour plates!
Dry lab
Discovered that the LVA tagging T7 RNA polymerase is a no go as the C-terminus is part of the active site of the enzyme. We'll have to do N-terminal tagging, this could work, see the crystal structure of T7. See the link under literature on this wiki. Lon-mediated degradation of lambda excisionase (Xis) and E.coli UmuD gene product. 30-40 amino temrinal AA of UmuD should do the trick.
Modeling
Parameters, parameters, parameters, ... and Matlab, aah... (total smiley face)
Discovered ETHZ 2007 parameter page revealing information on certain degradation and kinetical constants, the link is supplied on the Modeling page.
Meeting
Present: All students, IT, KM, JW, JR, BDM, AC
Some remarks we have got on our project:
- They showed us where we could find the right E.coli strains
- There could be a problem with photobleaching of eCFP
- The production of eCFP could also be lowered by the low efficiency of the RBS
- Will there be enough ribokey? (Hanne says: ENHANCER!)
- Enough lactonase?
- Maybe we should put tetR under control of his own promotor and lift out of the cell death construct
- We can inhibit our system exogenous by adding lactonase to our medium
- We should use different ori's for our different plasmids, perhaps we 'll have to link some devices
- The engineers should make a simple scheme that indicates which parameters are important for the sensitivity of our system
Remarks
How great it is to have a sandbox!
