Microbiology: An Introduction (13th Edition)
13th Edition
ISBN: 9780134605180
Author: Gerard J. Tortora, Berdell R. Funke, Christine L. Case, Derek Weber, Warner Bair
Publisher: PEARSON
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Textbook Question
Chapter 9, Problem 3A
The following picture shows bacterial colonies growing on X-gal plus ampicillin in a blue-white screening test. Which colonies have the recombinant plasmid? The small satellite colonies do not have the plasmid. Why did they start growing on the medium 48 hours after the larger colonies?
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Define the following terms:
Plasmid
Restriction Enzyme
Standard Curve
Explain how you would use the blue-white colony screening technique to identify bacteria containing the recombinant plasmid
Why are the following reagents used?
Neutralizing solution (Plasmid isolation)
Isopropanol (Plasmid isolation)
RNase (isolation of genomic DNA)
Chapter 9 Solutions
Microbiology: An Introduction (13th Edition)
Ch. 9 - Compare and contrast the following terms: a. cDNA...Ch. 9 - Differentiate the following terms. Which one is...Ch. 9 - Some commonly used restriction enzymes are listed...Ch. 9 - Suppose you want multiple copies of a gene you...Ch. 9 - Which enzyme makes the smallest fragment...Ch. 9 - Describe a recombinant DNA experiment in two or...Ch. 9 - List at least two examples of the use of rDNA in...Ch. 9 - You are attempting to insert a gene for saltwater...Ch. 9 - How does RNAi silence a gene?Ch. 9 - Prob. 10R
Ch. 9 - Restriction enzymes were first discovered with the...Ch. 9 - The DNA probe, 3-GGCTTA, will hybridize with which...Ch. 9 - Which of the following is the fourth basic step to...Ch. 9 - The following enzymes are used to make cDNA. What...Ch. 9 - If you put a gene in a virus, the next step in...Ch. 9 - You have a small gene that you want replicated by...Ch. 9 - Pieces of human DNA stored in yeast cells. a....Ch. 9 - A population of cells carrying a desired plasmid....Ch. 9 - Self-replicating DNA for transmitting a gene from...Ch. 9 - A gene that hybridizes with mRNA. a. antisense b....Ch. 9 - Design an experiment using vaccinia virus to make...Ch. 9 - Why did the use of DNA polymerase from the...Ch. 9 - The following picture shows bacterial colonies...Ch. 9 - Prob. 1CAECh. 9 - Using the restriction enzyme ECORI, the following...
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- When a 5-kb circular plasmid is digested with a restriction enzyme that has three recognition sites on the plasmid, how many bands can be visualized on an agarose gel? a)1 b)2 c)5 d)3arrow_forwardConstruct a restriction map for Plasmid X to show the recognition sites for two commonly used restriction enzymes: EcoRI and BamHI. Plasmid X undigested Plasmid X digested with EcoRI Plasmid X digested with BamHI Plasmid X digested with EcoRI and BamHI 2200 bp 1000 bp 1200 bp 400 bp 1800 bp 200 bp 400 bp 600 bp 1000 bparrow_forwardTo make the recombinant plasmid permeable to DNA molecules, which of the chemicals is added?a) MgCl2b) CaCl2c) NaCld) HClarrow_forward
- Please answer the following using the experiment data below. Provide the formulas or methods used for calculations of each. Number of colonies on LB/amp/ara plate = Micrograms of DNA spread on the plates = Transformation efficiency = DNA plasmid concentration: 0.08 μg/μl 250 μl CaCl transformation solution 10 μl pGLO plasmid solution 250 μl LB broth 100 μl cells spread on agar 227 colonies of transformantsarrow_forwardThe following image is of an agarose gel. If DNA samples were loaded to this gel and the electrophoresis experiment was started, explain what would happen and why.arrow_forwardIf the cloning was successful and the plasmid contained the elements needed for blue-white screening, what should be the color of the colonies harboring the correct plasmid?arrow_forward
- Why are there strict reminders about mixing in the inverted position and not vortexing Plasmid samples? Will these actions affect the plamid DNA extraction?arrow_forwardWhy are plasmids used as vector for DNA Recombination? What other vectors can be used?arrow_forwardChoose the one answer that fits best. The pGLO plasmid you used contains an araC gene, a GFP gene and a bla gene. If you performed the exact same experiment with a plasmid that contained a GFP gene and a bla gene, but was missing the araC gene, what would you expect to see on your four plates? a. The positive control would not have any bacterial lawn b. The negative control would have as many colonies as the experimental plate c. The LB/Amp/Ara plate would have some colonies but they would not be fluorescent under UV light d. The experimental plate would not have any colonies e. The LB/Amp plate containing E. coli (+)pGLO would be fluorescentarrow_forward
- When making recombinant DNA, why must you use the same restriction enzyme to cut the gene of interest and the plasmid?arrow_forwardYou have isolated genomic DNA and plasmid DNA from a bacterial culture. Please answer the following questions: 1. What solution did you use to elute DNA from the silica resin column? 2. What is the function of the chaotropic salt in DNA isolation? 3. Why do we add the chaotropic salt for genomic DNA before lysis and for plasmid isolation we add the salts after the lysis step?arrow_forwardA small DNA molecule was cleaved with several different restriction nucleases, and the size of each fragment was determined by gel electrophoresis. The following data were obtained.arrow_forward
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