1. Grow cells O/N at 30˚C in 200µl YPD
   
2. Spin down and wash in 200µl ddH2O
   
3. Spin down and wash in 200µl 0.1M LiOAc twice
   
4. Resuspend in 25µl 0.1M LiOAc and mix in 30µl ddH2O
   
5. Mix well with 120µl 50% PEG
   
6. Mix in 18µl 1M LiOAc + 2.5µl ssDNA + 4.5 µl plasmid (approximately 500ng)
   
7. Incubate O/N at 30˚C
   
8. Incubate 20 min at 42˚C
   
9. 1. Spin down and resuspend in 200µl drop-out medium
      Colonies should appear after 2-4 days at the bottom of each well
      or
   2. Spin down and resuspend in 10 µl ddH2O, streak-plate on solid drop-out medium. Colonies should appear after 2-4 days
      
      To save plates, the transformant mixes can be streaked in 8 lines across each drop-out plate using an 8-channel pipette.
      

Michael Lisby (December 2001)