Transgenesis Kit for the yeast S. cerevisiae and detection of GFP by PCR

Topics covered:
- Learning to work in sterile conditions in a microbiology laboratory
- Performing inter-species gene transfer
- Measuring GFP protein expression on selective medium
- Carrying out a complete PCR protocol

Transgenesis Lab
Principles and objectives: Jeulin has designed a protocol that does not require making cells competent. It simply involves resuspending fresh yeast cells from a colony, mixing with the transformation solution and plasmid DNA, vortexing, then incubating for 30 minutes at 42°C. The yeast cells are then cultured to verify the efficiency of the transfer, and the expression of the bioluminescence gene is revealed by illuminating the boxes with UV radiation.
This experiment lasts approximately 120 minutes + 1 results reading session
Composition: Yeast strain on a plate, ready-to-pour YPD and URA- medium, Petri dishes, spreaders, sterile sticks, sterile microtubes

PCR Lab
Principles and objectives: Verify the presence of the GFP gene responsible for the fluorescence of a transformed yeast through PCR amplification.
Experiment in 3 steps: sample extraction/purification, PCR amplification, and agarose gel electrophoresis.
Students can also test the strains obtained through the Transgenesis Lab. The protocol and primers are compatible.
Plan for 2 sessions of 60 minutes for this experiment. The kit is designed for 20 PCRs.
Composition: PCR microtubes, primers, ready-to-use PCR reagents, 2 yeast strains, molecular weight ladder

Comments: We offer two kits that can be done separately or one after the other for a complete experimentation linking proteomics and genomics in a microbiology laboratory.