Maheshri:TransformationY

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A 'sleazy' method for plasmid transformation

1) Using sterile technique, mix the following in a microcentrifuge tube:

PEG 3500 50% W/V        240μL
LiAc 1.0M               36μL
Boiled SS-carrier DNA   50μL
Plasmid DNA plus ddH2O  34μL
A toothpick full of yeast from a fresh plate (<2 days old)

2) Incubate the tubes at room temperature for overnight (>8 hours)

3) Microcentrifuge at 3000rpm for 1 min and remove the Transformation Mix

4) Optional - Wash (resuspend and pellet) the cells with 1.0 mL of sterile water

5) Add 200μL of sterile water to the tube, resuspend cells, and plate the cell suspension onto selection medium (e.g. SD dropout plates).