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  1. Preparation of materials
  • Agar plates w/ antibiotics
  • Agarose gels
  • Competent cells


  1. Experimentation
  • Miniprep/Midiprep
  • Cloning of GFP into pUC19 (digestion, PCR, gel excision, ligation, transformation, colony picking)
  • Cloning of sgRNA into PX459 (digestion, annealing oligos, ligation, transformation, colony picking)
  • Verification of cloning by restriction digest
  • Colony PCR
  • RNA extraction
  • Reverse transcription
  • qPCR
  1. Analysis
  • Primer selection
  • Enzyme selection
  • Clonings
  • qPCR results

Compétences visées

  • Understanding the protocols used in genetic engineering
  • Acquiring the theoretical bases of cloning
  • Acquiring a good practice in genetic engineering
  • Being able to analyze and discuss experimental results
  • Writting of a lab book
  • Cleaning of the room and machines used



Bibliography, course material : Protocols and documents necessary to perform the experiments are given before the practical and all documents are available on Moodle  


Responsable(s) de l'enseignement
Anne-Laure Duchemin :