"
Team:Bielefeld-Germany/Project/Protocols
From 2010.igem.org
(Difference between revisions)
| Line 19: | Line 19: | ||
: Error Proune PCR, DNA shuffling, directed evolution, Protein coupling assay | : Error Proune PCR, DNA shuffling, directed evolution, Protein coupling assay | ||
| - | * contact to local newspapers | + | * contact to local newspapers; TV; Radio |
| Line 48: | Line 48: | ||
* cloning of a constitutive promotor and a rbs in front of the improved virA | * cloning of a constitutive promotor and a rbs in front of the improved virA | ||
| + | * cloning of virG into the right biobrick vector | ||
| + | * creating new antibiotic biobricks | ||
| + | |||
| + | * Testing Top10 and Ec100D for ccdb-gene | ||
=== to be done === | === to be done === | ||
| - | |||
* characterisation of new build standalone virG biobrick | * characterisation of new build standalone virG biobrick | ||
* characterisation of new build virA biobrick | * characterisation of new build virA biobrick | ||
| - | |||
| - | |||
* cloning of the construct backbone (promotor, virA, terminator, virB, virG, readout) | * cloning of the construct backbone (promotor, virA, terminator, virB, virG, readout) | ||
Revision as of 08:47, 23 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; TV; Radio
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
- cloning of virG into the right biobrick vector
- creating new antibiotic biobricks
- Testing Top10 and Ec100D for ccdb-gene
to be done
- characterisation of new build standalone virG biobrick
- characterisation of new build virA biobrick
- cloning of the construct backbone (promotor, virA, terminator, virB, virG, readout)
- Error Proune PCR of virA
- Sensitivity test by antibiotic gradient
- Modelling
- Constructing a new Backboneplasmid -> creating a cloning system in order to insert the r6k-origin in all psb-backbone plasmids
- Testing of the ccdb-death gene
