Objectives in terms of knowledge (course content) :

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

 

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

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