This protocol was submitted by Matt Kaeberlein . Streak yeast onto YPD plates and incubate overnight at 30C. Restreak onto fresh YPD the next day and incubate overnight at 30C. Patch onto fresh YPD early on 3rd day. Incubate at 30C for 2-3 hours.
Some people do one strain per plate.
Dip end of micromanipulator into patch of cells. Move 50-100 cells about half way across plate from patch and deposit cells on agar. Incubate at 30C for 1.5-3 hours. Each of the cells you positioned in step 5 should divide one or two times. This is the only time you will harvest daughter cells.