Prescott's Microbiology
10th Edition
ISBN: 9781259281594
Author: Joanne Willey, Linda Sherwood Adjunt Professor Lecturer, Christopher J. Woolverton Professor
Publisher: McGraw-Hill Education
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Chapter 28.1, Problem 2.2RIA
Summary Introduction
All living organisms (plants and animals) require nitrogen. Nitrogen is a chemical element used to generate proteins. Ammonia is a type of nitrogen present in soil and it enters the soil in this organic form from dead and decaying animals and plants waste. Plants unable to absorbs the organic form of nitrogen. Hence, the organic form of nitrogen must be converted into inorganic form, through a process called nitrification.
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1) Look at the ideal results. Were your predictions accurate, and how did they compare with your results?
2) You used aseptic technique during this lab. Why is it important to work in a sterile manner when working with bacteria in the lab?
3) Why are the cells incubated at 42°C?
Overview of Transformation Protocol
-Prepare competent bacteria for transformation:
Treat starter E. coli bacteria with CaCl2and Competent Cell Solution (CCS). Store on ice until transformation procedure.
Competent cells are cells that are likely to take up foreign DNA and be transformed. This step increases the likelihood that the E. coli cells will take up the introduced vector and be transformed.
-Transformation procedure:
Obtain two microcentrifuge tubes containing your competent cells. Label one tube +DNA and one -DNA.
Add CaCl2 to both tubes.
Add the transformation mix containing the plasmid DNA to the tube labeled +DNA. Do not add any plasmid DNA to the -DNA tube.
Incubate both tubes on ice for 10 minutes. Then, place both tubes in a 42\deg C water bath for 45 seconds. Replace the tubes in an ice bucket for 2 minutes.
Add recovery broth to both tubes.
Incubate both tubes in a 37 C water bath for 5 minutes.
Questions:
1) What differences would you expect to see between the…
Overview of Transformation Protocol
-Prepare competent bacteria for transformation:
Treat starter E. coli bacteria with CaCl2and Competent Cell Solution (CCS). Store on ice until transformation procedure.
Competent cells are cells that are likely to take up foreign DNA and be transformed. This step increases the likelihood that the E. coli cells will take up the introduced vector and be transformed.
-Transformation procedure:
Obtain two microcentrifuge tubes containing your competent cells. Label one tube +DNA and one -DNA.
Add CaCl2 to both tubes.
Add the transformation mix containing the plasmid DNA to the tube labeled +DNA. Do not add any plasmid DNA to the -DNA tube.
Incubate both tubes on ice for 10 minutes. Then, place both tubes in a 42\deg C water bath for 45 seconds. Replace the tubes in an ice bucket for 2 minutes.
Add recovery broth to both tubes.
Incubate both tubes in a 37 C water bath for 5 minutes.
Questions:
1)What is the selectable marker in this experiment? How…
Chapter 28 Solutions
Prescott's Microbiology
Ch. 28.1 - Prob. 1MICh. 28.1 - What microbes are capable of methanogenesis and...Ch. 28.1 - What are the products of denitrification,...Ch. 28.1 - Prob. 4MICh. 28.1 - Prob. 5MICh. 28.1 - Prob. 1.1RIACh. 28.1 - Prob. 1.2RIACh. 28.1 - Prob. 1.3RIACh. 28.1 - Prob. 1.4RIACh. 28.1 - Prob. 1.5RIA
Ch. 28.1 - Prob. 2.1RIACh. 28.1 - Prob. 2.2RIACh. 28.1 - Prob. 2.3RIACh. 28.1 - Prob. 2.4RIACh. 28.1 - Compare the products of dissimilatory sulfate...Ch. 28.1 - Prob. 3.2RIACh. 28.1 - Prob. 3.3RIACh. 28.1 - Prob. 3.4RIACh. 28.1 - Suggest how a chemolithoautotroph might link the...Ch. 28.2 - What organisms benefit from nitrification?Ch. 28.2 - List three greenhouse gases. Discuss their...Ch. 28.2 - Prob. 2RIACh. 28.2 - How do changes in the nitrogen cycle caused by...Ch. 28.2 - Prob. 4RIACh. 28 - Compare the production of N2 gas by...Ch. 28 - Prob. 2CHICh. 28 - A bacterium isolated from sewage sludge was...Ch. 28 - It is well documented that aquatic ecosystems have...
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