Control #1: E.
coli in the presence of the DNA plasmid is cultured on LB Agar with no
ampicillin antibiotic.
Is this positive or negative control?
This is positive control, because the [Link] was still in the dish after transformation.
What do you expect [Link] to do on this plate?
I expect [Link] to grow on this plate.
Actual Results:
There were a few colonies of bacteria clustered on one side of my LB+ plate.
Control #2: [Link] goes through the transformation procedures in the absence of the
plasmid and is cultured on the LB agar plate.
Is this positive or negative control?
This is positive control, because the [Link] was still in the dish after transformation.
What do you expect [Link] to do on this plate?
I expect [Link] to grow on this plate.
Actual Results:
There were many colonies of bacteria in a horizontal strip across my plate. They did not
form a lawn, because I could see each individual colony.
Control #3: [Link] goes through the transformation procedures in the absence of the
plasmid and is cultured on the LB/Amp agar plate.
Is this positive or negative control?
This is negative control, because the bacteria needed the DNA plasmid to be resistant to
ampicillin.
What do you expect [Link] to do on this plate?
I expect to have no bacteria on this plate, because the ampicillin would inhibit the
bacteria’s growth.
Actual Results:
I see no bacteria on this plate.
Experimental Plate: LB/Amp agar with E/coli transformed with plasmid.
What do you expect [Link] to do on this plate?
I expected a little [Link] to grow on this plate, because the DNA plasmid should have picked
up a little bacteria.
Actual results:
There was a very small amount of bacteria that was whiter than the colonies in other
plates.
Discussion Questions:
1. Compare and contrast the number of colonies on each of the following pairs of
plates. What does each pair of results tell you about the experiment?
● LB +Plasmid and LB -Plasmid
This pair told me that the bacteria cells were still in the plates after the
experiment.
● LB/Amp -Plasmid and LB -Plasmid
This pair told me that the ampicillin successfully prevented the growth of
bacteria cells that didn’t have plasmid DNA.
● LB/Amp +Plasmid and LB/Amp -Plasmid
This pair told me that some of the bacterial cells on the LB/Amp +Plasmid
plates became ampicillin resistant. They formed colonies on my plate.
● LB/Amp +Plasmid and LB +Plasmid
This pair told me that a small amount of bacteria took up the plasmid DNA.
The LB+Plasmid formed more colonies than the LB/Amp+Plasmid, although I
could still see each individual colony.
2. What are you selecting for in this experiment?
I am selecting for bacteria that are resistant to ampicillin.
3. What does the phenotype of the transformed colonies tell you?
The phenotype of the transformed colonies told me about which plates had bacteria
that were resistant to ampicillin.
4. What one plate would you first inspect to conclude that the transformation occurred
successfully? Why?
I would first inspect the LB/Amp+Plasmid plate, because it would only show the
bacterial colonies that were resistant to the antibiotic.
5. What is your transformation efficiency? To answer this question proceed to Lab
Investigation 7.2 to determine your transformation efficiency.
Transformation efficiency = (total number of colonies growing on LB/Amp
plate)/(amount of DNA spread on the LB/Amp plate) = 3colonies/250µl = 0.012