If you incubate your cultures at 30xC, you will be able to follow this schedule. At room temperature, you may have to double the incubation time.
1st Day : 10 min Streak Transformants
3rd Day: 15 min Count Red and Sectored Colonies
Map of Plasmid YCpADE1
Map of Plasmid YEpADE1
Time Line: 1st Day 1. Isolate single colonies from one of the transformant colonies. Touch a sterile toothpick to one of the white transformant colonies. Pick a large white colony (avoid tiny colonies). Make a streak on a new YED plate (See Figure 1).
Then use a new sterile toothpick to make another
zigzag streak across the first one on your YED
Continue using fresh sterile toothpicks to make 4 or 5 more zigzag streaks in this manner. The last streaks should give you some single colonies. Each colony will grow from a single transformant cell.
2. Follow the same procedure to streak
out some of the transformant cells on
an MV plate. (See Figure 1)
Time Line: 3rd Day
3. Count the total number of single
colonies on each plate.
Count the number of white colonies on each plate.
Use these numbers to compute the percent of
transformants in each plate.
4. Which condition maintained the
highest percentage of transformants?
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Last updated Tuesday November 18 1997