Team:Bielefeld-Germany/Project/Protocols

From 2010.igem.org

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* Modelling
* Modelling
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* Constructing a new Backboneplasmid -> creating a cloning system in order to insert the r6k-origin in all psb-backbone plasmids
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* Testing of the ccdb-death gene

Revision as of 12:11, 13 August 2010

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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
  • 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