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Plating Yeast Colonies 1. Swirl 1 colony (0.5 -2 mm diameter) into 1 ml sterile dH2O with toothpick (faintly turbid).
 

Summary: 78
Plating Yeast Colonies
1. Swirl 1 colony (0.5 - 2 mm diameter) into 1 ml sterile dH2O with toothpick (faintly turbid).
2. Dilute 2 - 10 Ál (depending on colony diameter) into 1 ml sterile ddH2O. Vortex.
3. Spread 100 Ál. For ~ 1 mm colony will yield roughly 500 - 1000 colonies.
Replica Plating with Pin Stamp
1. The pin stamp has 48 pins. The top left pin is a half pin for orienting replicas. Label a sterile 96-
well plate. Pipette 180 Ál of sterile ddH2O into each well. Use 6 rows of pins in the middle of the
stamp for best results.
2. Serial dilutions: Do 10-fold dilution series, such that each well has 10-fold fewer cells than the
previous well. Usually, 5 serial 10-fold dilutions is enough, but it is convenient to include a 6th
final well. Innoculate the right-hand well with a colony using a sterile toothpick (twirl toothpick)
or with 20 Ál liquid culture. Transfer 20 Ál to next well, repipette to mix. Repeat for all wells.
Undiluted Cells (2nd column from left is best)
Yeast Strain #1
Yeast Strain #2
Yeast Strain #3
Yeast Strain #4
Yeast Strain #5
Yeast Strain #6

  

Source: Aris, John P. - Department of Anatomy and Cell Biology, University of Florida

 

Collections: Biology and Medicine