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Team:EPF-Lausanne/Todo
From 2011.igem.org
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* <s> Research plasmid backbones. Is there one that already contains Tet-repressed LacI or GFP? </s> | * <s> Research plasmid backbones. Is there one that already contains Tet-repressed LacI or GFP? </s> | ||
* <s>Design Gibson primers to assemble the three different plasmids.</s> | * <s>Design Gibson primers to assemble the three different plasmids.</s> | ||
| - | * Receive said primers | + | * <s>Receive said primers</s> |
* Determine which sequence on LacI-plasmid and Lysis-plasmid should be used to create the "mega-plasmid". | * Determine which sequence on LacI-plasmid and Lysis-plasmid should be used to create the "mega-plasmid". | ||
* Think of new assemblies we want to make (pTet with RFP, for example) | * Think of new assemblies we want to make (pTet with RFP, for example) | ||
| Line 27: | Line 27: | ||
* determine required sequences | * determine required sequences | ||
* order primers | * order primers | ||
| - | * ext. PCR for MITOMI | + | * ext. PCR for MITOMI |
== MITOMI == | == MITOMI == | ||
Revision as of 08:32, 29 June 2011
Todo
Contents |
Microfluidic Chips
- Continue alignment training
- Run experiments on "chemostat" chip to check design
- Adapt design of "chemostat" chip for e-coli
Preparing the parts
-
Sequence the lysis cassette -
Double-check lysis cassette sequence
All the parts are verified, we can now assemble them!
Assembly
-
Research plasmid backbones. Is there one that already contains Tet-repressed LacI or GFP? -
Design Gibson primers to assemble the three different plasmids. -
Receive said primers - Determine which sequence on LacI-plasmid and Lysis-plasmid should be used to create the "mega-plasmid".
- Think of new assemblies we want to make (pTet with RFP, for example)
Specifically:
- Run PCRs and gels on existing sequences (plasmid backbone, TetR gene, RFP)
TetR mutants
- determine required sequences
- order primers
- ext. PCR for MITOMI
MITOMI
- repeat MITOMI experiments for practice
- order one-off sequences for tetR MITOMI
- de Brujin experiment on TetR (will also yield PWM)
- determine position weight matrix for TetR, compare with de Brujin results
- determine position weight matrix for TetR mutants
Microfluidics and chemostat chip
- Continue alignment training
- Repeat experiments to check design
- Grow E. Coli from spotted arrays
- [No microfluidics] Setup a plate and test tween concentrations
- Determine growth rate as function of tween concentration (say 0.075% +- 0.7, as many increments as will fit on the chip)
Wiki
Protocols
-
Describe cell cultures in miniprep protocol - Write a new protocol!
- Upload "chemostat" protocols
General
- Write-up team presentation
- Upload our initial research about transcription factors
Clean room
- Order lab notebook
-
Order storage box
