Description

Objectives in terms of knowledge (course content) :

Preparation of materials

• Agar plates w/ antibiotics
• Agarose gels
• Competent cells

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

Analysis

• Primer selection
• Enzyme selection
• Clonings
• qPCR results 

Compétences visées

Objectives in terms of skills:

• 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

Bibliographie

Protocols and documents necessary to perform the experiments are given before the practical and all documents are available on Moodle.
 

Contacts

Responsable(s) de l'enseignement

• Coraline Rigouin
• Anne-Laure Duchemin