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3. What do you think happened? How do you explain this result? Your supervisor gives you an E.coli stain (strain PIP) with an F plasmid on which is coded resistance for ampicillin. Your E.coli strain (strain ZAZ) is F- for the plasmid, and you would really like your strain to have ampicillin resistance. To favour conjugation, you grow both strains together for 2h (in broth) and plate agar containing ampicillin to grow single colonies overnight. grow overnight Inoculate agar plate containing ampicilline (amp plates) ZAZ colonies are blue, PIP colonies are yellow. Grow ZAZ and PIP together for 2h Figure 1. Schematic representation of the experimental plan. In the morning, you find multiple colonies growing on your ampicillin resistant agar plates. Because you had mixed both strains in the same culture, you needed to be able to distinguish between which colony is a ZAZ or a PIP colony growing on your plate. Fortunately, your ZAZ strain grows blue on the plates, so they are easily identifiable. To make sure the conjugation was successful, you choose 3 blue colonies and grow them in broth for 4h, then you isolate the plasmid and digest it with an enzyme that linearizes the plasmid. You know the F plasmid is 5,000bp long, which you use as a control (C) along your chosen 3 colonies. You run the plasmid on a gel and obtain the following result: 5Kb Marker Colonies Figure 2. DNA gel of linearized plasmids isolated from experimental bacterial colonies. Marker is a standard measure of DNA fragment size. C= control plasmid. Experimental colonies are numbered 1 to 3. The control F plasmid comes out at the expected size of 5kb, but all the colonies have plasmid that are bigger and not all the same size.

3. What do you think happened? How do you explain this result? Your supervisor gives you an E.coli stain (strain PIP) with an F plasmid on which is coded resistance for ampicillin. Your E.coli strain (strain ZAZ) is F- for the plasmid, and you would really like your strain to have ampicillin resistance. To favour conjugation, you grow both strains together for 2h (in broth) and plate agar containing ampicillin to grow single colonies overnight. grow overnight Inoculate agar plate containing ampicilline (amp plates) ZAZ colonies are blue, PIP colonies are yellow. Grow ZAZ and PIP together for 2h Figure 1. Schematic representation of the experimental plan. In the morning, you find multiple colonies growing on your ampicillin resistant agar plates. Because you had mixed both strains in the same culture, you needed to be able to distinguish between which colony is a ZAZ or a PIP colony growing on your plate. Fortunately, your ZAZ strain grows blue on the plates, so they are easily identifiable. To make sure the conjugation was successful, you choose 3 blue colonies and grow them in broth for 4h, then you isolate the plasmid and digest it with an enzyme that linearizes the plasmid. You know the F plasmid is 5,000bp long, which you use as a control (C) along your chosen 3 colonies. You run the plasmid on a gel and obtain the following result: 5Kb Marker Colonies Figure 2. DNA gel of linearized plasmids isolated from experimental bacterial colonies. Marker is a standard measure of DNA fragment size. C= control plasmid. Experimental colonies are numbered 1 to 3. The control F plasmid comes out at the expected size of 5kb, but all the colonies have plasmid that are bigger and not all the same size.

Human Anatomy & Physiology (11th Edition)
Human Anatomy & Physiology (11th Edition)
11th Edition
ISBN:9780134580999
Author:Elaine N. Marieb, Katja N. Hoehn
Publisher:Elaine N. Marieb, Katja N. Hoehn
Chapter1: The Human Body: An Orientation
Section: Chapter Questions
Problem 1RQ: The correct sequence of levels forming the structural hierarchy is A. (a) organ, organ system,...
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Practice question #3 chap. 2 p. 145 3. What do you think happened? How do you explain this result? Your supervisor gives you an E.coli stain (strain PIP) with an F plasmid on which is coded resistance for ampicillin. Your E.coli strain (strain ZAZ) is F- for the plasmid, and you would really like your strain to have ampicillin resistance. To favour conjugation, you grow both strains together for 2h (in broth) and plate agar containing ampicillin to grow single colonies overnight. grow overnight Inoculate agar plate containing ampicilline (amp plates) ZAZ colonies are blue, PIP colonies are yellow. Grow ZAZ and PIP together for 2h Figure 1. Schematic representation of the experimental plan. In the morning, you find multiple colonies growing on your ampicillin resistant agar plates. Because you had mixed both strains in the same culture, you needed to be able to distinguish between which colony is a ZAZ or a PIP colony growing on your plate. Fortunately, your ZAZ strain grows blue on the plates, so they are easily identifiable. To make sure the conjugation was successful, you choose 3 blue colonies and grow them in broth for 4h, then you isolate the plasmid and digest it with an enzyme that linearizes the plasmid. You know the F plasmid is 5,000bp long, which you use as a control (C) along your chosen 3 colonies. You run the plasmid on a gel and obtain the following result: 5Kb Marker 1 2 Colonies Figure 2. DNA gel of linearized plasmids isolated from experimental bacterial colonies. Marker is a standard measure of DNA fragment size. C= control plasmid. Experimental colonies are numbered 1 to 3. The control F plasmid comes out at the expected size of 5kb, but all the colonies have plasmid that are bigger and not all the same size.
Transcribed Image Text:Practice question #3 chap. 2 p. 145 3. What do you think happened? How do you explain this result? Your supervisor gives you an E.coli stain (strain PIP) with an F plasmid on which is coded resistance for ampicillin. Your E.coli strain (strain ZAZ) is F- for the plasmid, and you would really like your strain to have ampicillin resistance. To favour conjugation, you grow both strains together for 2h (in broth) and plate agar containing ampicillin to grow single colonies overnight. grow overnight Inoculate agar plate containing ampicilline (amp plates) ZAZ colonies are blue, PIP colonies are yellow. Grow ZAZ and PIP together for 2h Figure 1. Schematic representation of the experimental plan. In the morning, you find multiple colonies growing on your ampicillin resistant agar plates. Because you had mixed both strains in the same culture, you needed to be able to distinguish between which colony is a ZAZ or a PIP colony growing on your plate. Fortunately, your ZAZ strain grows blue on the plates, so they are easily identifiable. To make sure the conjugation was successful, you choose 3 blue colonies and grow them in broth for 4h, then you isolate the plasmid and digest it with an enzyme that linearizes the plasmid. You know the F plasmid is 5,000bp long, which you use as a control (C) along your chosen 3 colonies. You run the plasmid on a gel and obtain the following result: 5Kb Marker 1 2 Colonies Figure 2. DNA gel of linearized plasmids isolated from experimental bacterial colonies. Marker is a standard measure of DNA fragment size. C= control plasmid. Experimental colonies are numbered 1 to 3. The control F plasmid comes out at the expected size of 5kb, but all the colonies have plasmid that are bigger and not all the same size.
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