Essentials of Genetics (9th Edition) - Standalone book
Essentials of Genetics (9th Edition) - Standalone book
9th Edition
ISBN: 9780134047799
Author: William S. Klug, Michael R. Cummings, Charlotte A. Spencer, Michael A. Palladino
Publisher: PEARSON
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Chapter 8, Problem 20PDQ

A phage-infected bacterial culture was subjected to a series of dilutions, and a plaque assay was performed in each case, with the following results. What conclusion can be drawn in the case of each dilution?

Dilution Factor Assay Results
(a) 104 All bacteria lysed
(b) 106 14 plaques
(c) 108 0 plaques
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Shown below is a set of cell culture plates for a plaque assay.  The assay was performed by preparing dilutions of a virus stock with the dilution factor for each prepared dilution listed below the sample.  0.5 mls of each dilution was added to confluent (fully covered) monolayers of cells on each plate.  The virus used for the assay had a titer of 2 x 1010 Plaque forming units (PFU) per ml. Only 1 set of the above plates would have a number of plaques on it that would be both easy to count and high enough to be statistically relevant.  Which dilution set would it be and about how many plaques would there be on the plates of that dilution set.
A phagehunter performs a spot titer using standard techniques (3 ul of each dilution spotted to lawn) of a lysate obtained from an optimum webbed plate experiment. The phagehunter counts 4 plaques on the 10-8 dilution spot. Which of the following scenarios is the best choice for the phagehunter to do next? a.) The phagehunter has not achieved a high enough titer lysate to move forward with characterization experiments, so they should try to make more optimum webbed plates. b.) The phagehunter has not achieved a high enough titer lysate to move forward with characterization experiments, so they should adopt a phage from direct isolation. c.) The phagehunter has achieved a high enough titer lysate to move forward with characterization experiments. d.) The phagehunter has not achieved a high enough titer lysate to move forward with characterization experiments, so they should go back to the pick-a-plaque experiment.
3) Were all of the conditions of a standardized Kirby-Bauer test met as you performed this assay? If not, which were not? 4) What is the significance of colonies that develop within otherwise clear zones of inhibition? If the laboratory report for one of your patients indicated colonies within the zone, what concerns would you have for your patient?

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