Team:Washington/Protocols/Yeast

From 2012.igem.org


Yeast Transformation

Yeast Transformation (for 6 transformations)

  • Combine 50uL of competent yeast EBY100 cells with:
    • 240uL 50% PEG
    • 36uL of 1 M LiOAc
    • 28uL of sterile dH2O to have a total volume of 354uL.
  • Aliquot 59uL of yeast mixture into 6 tubes.
  • Add 1uL of plasmid to each tube
  • Vortex hard to mix
  • Incubate at 30° C for 30 minutes
  • Heat shock at 42° C for 20 minutes
  • Plate 30uL on selective media.
    • we used C-Trp-Ur plates