Microbiology Fundamentals: A Clinical Approach
3rd Edition
ISBN: 9781260163698
Author: Cowan
Publisher: MCG
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Chapter 11.2, Problem 12AYP
Draw a diagram of the stages of disease in a human
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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 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…
Based on your results, which suspect's DNA best matches the DNA found at the crime scene?
Chapter 11 Solutions
Microbiology Fundamentals: A Clinical Approach
Ch. 11.1 - Prob. 1AYPCh. 11.1 - Enumerate the sites where normal biota is found in...Ch. 11.1 - Discuss how the Human Microbiome Project is...Ch. 11.1 - Prob. 2NPCh. 11.2 - Differentiate between a microbes pathogenicity and...Ch. 11.2 - Prob. 5AYPCh. 11.2 - Explain the significance of polymicrobial...Ch. 11.2 - Prob. 7AYPCh. 11.2 - Prob. 8AYPCh. 11.2 - Prob. 9AYP
Ch. 11.2 - Prob. 10AYPCh. 11.2 - Prob. 11AYPCh. 11.2 - Draw a diagram of the stages of disease in a humanCh. 11.2 - Prob. 13AYPCh. 11.2 - Prob. 14AYPCh. 11.2 - Define healthcare-associated infection, and list...Ch. 11.2 - Prob. 16AYPCh. 11.2 - Prob. 1MMCh. 11.2 - Prob. 3NPCh. 11.2 - Prob. 2MMCh. 11.2 - Prob. 4NPCh. 11.2 - Prob. 5NPCh. 11.2 - Q. Can you think of what type of incident might...Ch. 11.2 - Prob. 6NPCh. 11.3 - Summarize the goals of epidemiology, and...Ch. 11.3 - Prob. 18AYPCh. 11.3 - Prob. 19AYPCh. 11.3 - Discuss the three major types of epidemics, and...Ch. 11 - Prob. 1NPCh. 11 - Prob. 1QCh. 11 - Prob. 2QCh. 11 - Prob. 3QCh. 11 - Prob. 4QCh. 11 - Prob. 5QCh. 11 - Discuss the role of endospores in ensuring the...Ch. 11 - Prob. 7QCh. 11 - Correlate the stages in the course of an...Ch. 11 - Prob. 9QCh. 11 - Prob. 10QCh. 11 - Prob. 11QCh. 11 - Prob. 12QCh. 11 - Prob. 13QCh. 11 - Prob. 14QCh. 11 - Prob. 15QCh. 11 - Prob. 16QCh. 11 - Prob. 17QCh. 11 - Prob. 18QCh. 11 - The number of cases, including new cases as well...Ch. 11 - Prob. 20QCh. 11 - One important way to critically analyze a study...Ch. 11 - Prob. 1VC
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- In 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_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. 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_forward
- Why 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_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. 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_forward
- is often a good indication of phylogenetic relatedness in phenotypes. Life-cycle patterns Cleavage patterns O Gene expression O Morphological similarityarrow_forwardWhich 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_forward
- What 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_forwardWhich of the following sequencing techniques can identify down to the strain level? O Multilocus sequence typing Genomic fingerprinting Whole genome sequencing OSNP analysis All of the abovearrow_forwardWhat is the "gold standard" that is currently applied to species designations in microbiology? 97% between species: 50% among whole genome 90% between species: 75% among whole genome 99% between species; 97% among whole genome 97% between species: 70% among whole genome Onone of the abovearrow_forward
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