Transformations of Yeast

There are a variety of methods to transform yeast.

Chemical transformations is used most in the lab

• First make Synchronized Competent Yeast Cells (Takes 1 day prep + 1 day handling time)
• Freeze these to -80°C to increase competence
• Take aliquots of competent cells from the -80°C
• Transform the cells using (<1h + ON step):
  • Yeast Chemical Transformations for homologous recombination or;
  • CRISPR/Cas9 yeast transformations for CRISPR
• Plate out on selective media and incubate (~3-5 days)
• Test colonies for transformation success (~1 week)

This method is most convenient and rather successful in our hands.

Alternatively use electroporation

• First make Yeast Electoporation Competent cells
• Freeze these to -80°C to increase competence
• Take aliquots of competent cells from the -80°C
• Transform the cells using Yeast Electoporation Transformation (equipment in Gel Room)
• Plate out on selective media and incubate
• Test colonies for transformation success

In theory, this should yield higher transformation efficiency, however, this is in our experience not worth the effort of setting up the equipment.

Chemical transformations from yeast colonies

• First make Yeast cells competent using PLATE
• Transform the cells using Yeast Colony Transformations protocol
• Plate out on selective media and incubate
• Test colonies for transformation success

This method is convenient as it takes little prep time to generate competent cells, but has very low transformation efficiency.