Microbiology Fundamentals: A Clinical Approach
3rd Edition
ISBN: 9781260163698
Author: Cowan
Publisher: MCG
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Chapter 3.2, Problem 6AYP
Summary Introduction
Toexplain:
The way through which a flagellum works in the presence of an attractant.
Concept introduction:
The bacteria are the prokaryotes,which have no organelles. They are single-celled organisms. Their cell wall has peptidoglycan and they reproduce by binary fission. There are few accessory structures that are originated from the exterior of a bacterial cell. These structures are not always present in all bacteria. They are divided into two groups: those which provide motility and those which provide channels or attachment points.
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1)Which plate did you see purple/pink/blue bacterial cells? Why did you see this growth? Explain your answer in terms of transformation and plasmids?
2) Calculating Transformation Efficiency
For the +DNA/+Amp/+IPTG plate, record the following:
Number of transformants (colonies): _________________
Nanograms of plasmid DNA added: 50 ng
Final recovery volume: 0.50 mL
Volume plated: 0.25 mL
Transformation efficiency equation:
Transformation efficiency = Number of transformants / µg of DNA x Final volume at recovery (mL)/ volume plated (mL)
3) Using the equation above, calculate the transformation efficiency.
4) Describe the success of the transformation efficiency of this demo based on the calculation you did above?
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…
Chapter 3 Solutions
Microbiology Fundamentals: A Clinical Approach
Ch. 3.1 - List the structures all bacteria possess.Ch. 3.1 - Identify three structures some but not all...Ch. 3.1 - Describe three major shapes of bacteria.Ch. 3.1 - Provide at least four terms to describe bacterial...Ch. 3.2 - Describe the structure and function of six...Ch. 3.2 - Prob. 6AYPCh. 3.2 - Q. Device-associated infections are very common...Ch. 3.3 - Differentiate between the two main types of...Ch. 3.3 - Prob. 8AYPCh. 3.3 - Prob. 9AYP
Ch. 3.3 - Prob. 2MMCh. 3.4 - Identify seven structures that may be contained in...Ch. 3.4 - Prob. 11AYPCh. 3.4 - Prob. 1NPCh. 3.5 - Compare and contrast the major features of...Ch. 3.6 - Differentiate between Bergeys Manual of Systematic...Ch. 3.6 - Name four divisions ending in cutes and describe...Ch. 3.6 - Define a species in terms of bacteria.Ch. 3 - Archaea a. are most genetically related to...Ch. 3 - Prob. 2QCh. 3 - Suppose an argument in your city has erupted about...Ch. 3 - Prob. 4QCh. 3 - As a supervisor in the infection control unit, you...Ch. 3 - Prob. 6QCh. 3 - Prob. 7QCh. 3 - Prob. 8QCh. 3 - Bacteria and archaea have a much greater diversity...Ch. 3 - Prob. 10QCh. 3 - Bacteria have been found to change the structures...Ch. 3 - Bacterial and archaeal chromosomes are not...Ch. 3 - Prob. 13QCh. 3 - The results of your patients wound culture just...Ch. 3 - We know that bacteria/archaea and their genetics...Ch. 3 - Find the true statement about biofilms. a. They...Ch. 3 - Suggest more than one reason why bacteria may...Ch. 3 - Construct arguments agreeing with and refuting...Ch. 3 - Which of the following would be used to identify...Ch. 3 - During the cold war between the Soviet Union and...Ch. 3 - During the cold war between the Soviet Union and...Ch. 3 - From chapter 2, figure 2.18. Explain why some...
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- 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…arrow_forwardBased on your results, which suspect's DNA best matches the DNA found at the crime scene?arrow_forwardIn oxidase test with Pseudomonas aeruginosa, the cell cultures on the slide turn colorless to be purple after tetra-methyl-p-phenylenediamine dihydrochloride (TMPD) is added. In the reaction, OTMPD is electron acceptor O cytochrome c is the electron source oxygen is terminal electron acceptor OH2 produced is electron donorarrow_forward
- You will use the following scenario to answer a group of 5 questions. You have isolated a microbe from an environmental sample. The microbe has the ability to perform a new metabolic reaction at a very low temperature, so you are excited that it could be a new species. You have shipped your samples off for sequencing and are now waiting for the results. Out of curiosity (and maybe boredom...) you decide to test your culture for the Catalase and Oxidase enzymes. Upon testing your sample for catalase, you don't see any bubbles; however, you do see a color change to purple during the Oxidase test. What results can you conclude from this? O Catalase-/ Oxidase + O Catalase +/ Oxidase + Catalase + / Oxidase- O Catalase / Oxidase - O None of the abovearrow_forwardWhich of the following is not a strength of using 16S rRNA for phylogenetic analyses? OA. It's cheap OB. It's easy to do C. It can be used to identify all the way down to the strain level OD. Both A & B OE. None of the abovearrow_forwardWhy are molecular approaches important to the field of microbial taxonomy and phylogeny? Phylogenetic inferences based on molecular approaches provide the most robust analysis of microbial evolution currently available. It allows for the collection of a large and accurate dataset from many organisms Almost no fossil record was left by microbes when compared to plants and animals All of the above None of the abovearrow_forward
- You will use the following scenario to answer a group of 5 questions. You have isolated a microbe from an environmental sample. The microbe has the ability to perform a new metabolic reaction at a very low temperature, so you are excited that it could be a new species. You have already cultured it and gone through the plate isolation procedure. Before you ship your samples off for sequencing, you want to do one final check of the A260 ratios. You get back the following ratios: A260/280 ratio is 1.89; A260/230 is 2.01. These ratios are close enough to the accepted "pure" values so they could be considered "pure" and mostly (if not completely) free of contaminants from the PCR process. True Falsearrow_forwardYou will use the following scenario to answer a group of 5 questions. You have isolated a microbe from an environmental sample. The microbe has the ability to perform a new metabolic reaction at a very low temperature, so you are excited that it could be a new species. After receiving your sequence back from the sequencing lab, you feel that you have, in fact, discovered and isolated a new species. You ask a fellow labmate about how you should proceed, and he tells you the following is the proper way to introduce a new species for recognition: Cultures have to be sent to international culture collections. Then a paper must be published describing the new organism and providing a genus and species name. You recall learning about this in your Microbiology course in college. Is this information from your colleague true or false? True Falsearrow_forwardis often a good indication of phylogenetic relatedness in phenotypes. Life-cycle patterns Cleavage patterns O Gene expression O Morphological similarityarrow_forward
- Which of the following is a weakness of using 16S rRNA for phylogenetic analyses? It can only go down to the family and genus levels It takes months to complete O Both of the above O None of the abovearrow_forwardAn unrooted tree containing ten unrelated species can become rooted by adding a descendant group related to two of the species. an unrelated outgroup. O a distantly related outgroup. O a descendant related to only one of the species.arrow_forwardWhat is the most appropriate purpose of building a phylogenetic tree? They look awesome You can use a tree to compare morphological characteristics of organisms It can be used to establish and analyze evolutionary relationships between species All of the abovearrow_forward
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