Strain, reagents and pouring medium (3 bottles of 350 ml)
The Earth in the universe, life and the evolution of life
Cognitive objective:
Transgenesis shows that genetic information is contained in the DNA molecule, and that it is written there in a universal language.

The Ade2 strain is :

Unable to grow on adenine-free medium.
Colored red on rich medium (YPD2).
This phenotype is due to a mutation in the adenine biosynthesis pathway.

Principle :
After introduction of a plasmid DNA fragment carrying the wild-type allele of the Ade2 gene into the Ade2 - strain, the latter will be able to grow on adenine-free medium and its staining on adenine-free medium will turn white.

Composition :

STRAINS, reagents and casting media (ref. TRAN/SMAC):

• Saccharomyces cerevisae Ade2 on Petri dish
• MEDIA and reagents :
  • 1 liter of selective medium
  • 100 ml rich medium for strain subculture
  • 50 µl plasmid
  • 20 ml lithium buffer
  • 5 ml transformation buffer
  • 3 tubes of sterile water

Equipment (for 40 students): optional (ref. TRAN/MAT)

• 50 Petri dishes, diameter 90 mm
• 40 color-coded microtubes with conical bottoms
• 20 inoculators
• 50 sterile spreaders
• 4 sterile bulbs

Technical and teaching guide: online


Equipment required :

• Centrifuge (3,000 rpm)
• Water bath (42 °C)
• Micropipettes

Storage: 2 months
Swabs and strain: refrigerator. Beware: the strain turns white in the fridge, so you'll need to re-transplant it if you keep it for more than two weeks.
Plasmid: freezer