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Team:Bielefeld-Germany/Project/Protocols
From 2010.igem.org
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(New page: {{Bielefeld_MainMenu_2010}} ==== Organisation and logistic ==== - flights in the US - Discussion of possible Substances for detection :-> list of Substances: :2-Chlorphenol (drug), Cap...) |
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| - | == | + | == '''Organisation and logistic''' == |
| - | + | * flights in the US | |
| - | + | * Discussion of possible Substances for detection | |
| - | : | + | :* list of Substances: |
| - | :2-Chlorphenol (drug), Capcaicin (spiciness), Dopamin and its derivates (human hormones), | + | ::2-Chlorphenol (drug), Capcaicin (spiciness), Dopamin and its derivates (human hormones), |
| - | :2,4,6 Trichloranisol(Responsible for bad taste in red wine)) | + | ::2,4,6 Trichloranisol(Responsible for bad taste in red wine)) |
| - | + | * Literature research for the virA sensor system | |
:contact of reaserch groups in order to get an already working system (failed) | :contact of reaserch groups in order to get an already working system (failed) | ||
| - | + | * Evaluation of the mutageneses strategy | |
| - | : | + | : Error Proune PCR, DNA shuffling, directed evolution, Protein coupling assay |
| - | + | * contact to local newspapers | |
| - | == | + | == '''Wetlab''' == |
| - | + | === accomplished === | |
| - | + | * qRT-PCR of induced ''aggrobacterium tumefaciens c58'' | |
| - | + | ||
:-> the strain could be significantly induced by acetosyringon | :-> the strain could be significantly induced by acetosyringon | ||
| - | + | * Synthesis of the virG by MrGene (use without rpoA, clear of illegal restriction sites, codon usage for ''a.tumrfaciens'') | |
| - | + | * Testing of the Promega readout machine (GloMax multiplate reader) for the LUC-assay (working) | |
:-> calibration of the GloMax | :-> calibration of the GloMax | ||
| - | + | * Testing of the virA construct from another researchgroup | |
:-> the virA construct could not be amplified | :-> the virA construct could not be amplified | ||
| - | + | * Testing of the virA biobrick taken of the iGEM regestry | |
:-> Correction and improvement of the virA biobrick | :-> Correction and improvement of the virA biobrick | ||
| - | + | * cloning of a constitutive promotor and a rbs in front of the improved virA | |
| - | + | === to be done === | |
| - | + | * cloning of virG into the right biobrick vector | |
| - | + | * characterisation of new build standalone virG biobrick | |
| - | + | * characterisation of new build virA biobrick | |
| - | + | * cloning of the antibiotic resistence in the correct biobrick backbone | |
| - | + | * cloning of the construct backbone (promotor, virA, terminator, virB, virG, readout) | |
| - | + | * Error Proune PCR of virA | |
| - | + | * Sensitivity test by antibiotic gradient | |
| - | + | * Modelling | |
Revision as of 19:04, 10 August 2010
{{{1}}}
Contents |
Organisation and logistic
- flights in the US
- Discussion of possible Substances for detection
- list of Substances:
- 2-Chlorphenol (drug), Capcaicin (spiciness), Dopamin and its derivates (human hormones),
- 2,4,6 Trichloranisol(Responsible for bad taste in red wine))
- Literature research for the virA sensor system
- contact of reaserch groups in order to get an already working system (failed)
- Evaluation of the mutageneses strategy
- Error Proune PCR, DNA shuffling, directed evolution, Protein coupling assay
- contact to local newspapers
Wetlab
accomplished
- qRT-PCR of induced aggrobacterium tumefaciens c58
- -> the strain could be significantly induced by acetosyringon
- Synthesis of the virG by MrGene (use without rpoA, clear of illegal restriction sites, codon usage for a.tumrfaciens)
- Testing of the Promega readout machine (GloMax multiplate reader) for the LUC-assay (working)
- -> calibration of the GloMax
- Testing of the virA construct from another researchgroup
- -> the virA construct could not be amplified
- Testing of the virA biobrick taken of the iGEM regestry
- -> Correction and improvement of the virA biobrick
- cloning of a constitutive promotor and a rbs in front of the improved virA
to be done
- cloning of virG into the right biobrick vector
- characterisation of new build standalone virG biobrick
- characterisation of new build virA biobrick
- cloning of the antibiotic resistence in the correct biobrick backbone
- cloning of the construct backbone (promotor, virA, terminator, virB, virG, readout)
- Error Proune PCR of virA
- Sensitivity test by antibiotic gradient
- Modelling
