Biology: The Dynamic Science (MindTap Course List)
Biology: The Dynamic Science (MindTap Course List)
4th Edition
ISBN: 9781305389892
Author: Peter J. Russell, Paul E. Hertz, Beverly McMillan
Publisher: Cengage Learning
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Chapter 17, Problem 14TYK
Summary Introduction

To review:

The experiment that can confirm the presence of a wild type bio+ allele in the plasmid of chromosomal DNA (deoxyribonucleic acid).

Introduction:

Minimal media is the media that only provides minimum requirement that is only inorganic salts and no amino acids. The colonies whose genome is complete, that is, they have genes for all requirements grows on the minimal media while others do not.

Electroporation is the technique in which electric pulse is given to the culture leading the opening of the transient pores in the cells that can allow the entry of the DNA from the surrounding.

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After transformation you were asked to grow bacterial cells transformed with plasmid on a plate that had X-gal and ampicillin. X-Gal is often used as in indicator dye, which turns blue when metabolized by B-galactosidase protein and used to test if cloning experiments have worked. [Note look at the vector diagrams carefully] Briefly explain how you would find the bacterial cells that are transformed with the plasmid with the YFG inserted.
you have transformed E.coli cells with a plasmid containing the gene for the enzyme beta-lactamase you have used 3µl of a stock solution of 2.5ng/µl DNA. You transformed 150/µl of competent cells, using the calcium chloride method. you have added 600µl of luria broth to the cells and then plated out 200µl on triplicate plates. upon counting the colonies, you got the following results: plate 1: 120 cfu plate 2: 35 cfu plate 3: 95 cfu please answer the following question: 1. what new characteristic will the cells have ater transformation? 2. how are you going to test for the new characteristic? 3. calculate the transformation efficiency of the experiment?
You have just carried out a transformation using a plasmid (possessing a Amp-resistance gene) that was 0.001 ug/ul in concentration. You added 10 ul of this plasmid to 100 ul of a bacterial cell suspension. Then you added 250 ul of LB following heat shock. After plating 200 ul of this cell suspension with LB onto a LBA plate, you observe 8 colonies on this plate. What is the transformation efficiency of this plasmid (in colonies/ug of plasmid) based on these results? about 1400 O about 80O about 4000 O about 400O O about 8000 f6 米 IO fs fg f1o 19 08. O OO
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