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 13.7, Problem 4RIA
Retrieve, Infer, Apply
Tabulate the nature and function of the following: fMet-tRNA, start codon, initiation factors, elongation cycle, elongation factors, peptidyl and aminoacyl sites, transpeptidation reaction, peptidyl transferase, translocation, stop codon, and release factors.
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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 13 Solutions
Prescott's Microbiology
Ch. 13.1 - MICRO INQUIRY Based on what we now know about...Ch. 13.1 - Retrieve, Infer, Apply 1. Briefly summarize the...Ch. 13.1 - Retrieve, Infer, Apply 2. Explain how protein was...Ch. 13.2 - MICRO INQUIRY To which carbon of ribose...Ch. 13.2 - MICRO INQUIRY How many H bonds are there between...Ch. 13.2 - Prob. 3MICh. 13.2 - Prob. 1RIACh. 13.2 - Retrieve, Infer, Apply What does it mean to say...Ch. 13.2 - Retrieve, Infer, Apply Amino acids are described...Ch. 13.3 - MICRO INQUIRY What provides the energy to fuel...
Ch. 13.3 - MICRO INQUIRY What is the difference between...Ch. 13.3 - MICRO INQUIRY Why cant DNA polymerase I perform...Ch. 13.3 - Retrieve, Infer, Apply How many replicons do...Ch. 13.3 - Retrieve, Infer, Apply Describe the nature and...Ch. 13.3 - Retrieve, Infer, Apply Outline the steps Involved...Ch. 13.3 - Retrieve, Infer, Apply What is the end replication...Ch. 13.4 - Why is the nontemplate strand called the sense...Ch. 13.4 - Retrieve, Infer, Apply The coding region of a gene...Ch. 13.4 - Which strand of a gene has sequences that...Ch. 13.4 - Briefly discuss the general organization of tRNA...Ch. 13.5 - MICRO INQUIRY Are the -35 and -10 regions...Ch. 13.5 - Retrieve, Infer, Apply Outline the transcription...Ch. 13.5 - Retrieve, Infer, Apply What is a polycistronic...Ch. 13.5 - Retrieve, Infer, Apply What is a consensus...Ch. 13.5 - Tabulate the similarities and differences between...Ch. 13.6 - Prob. 1MICh. 13.6 - Retrieve, Infer, Apply List the punctuation codons...Ch. 13.6 - What is the difference between a codon and an...Ch. 13.6 - Retrieve, Infer, Apply What is meant by code...Ch. 13.6 - Retrieve, Infer, Apply Is the genetic code truly...Ch. 13.7 - MICRO INQUIRY Why is simultaneous transcription...Ch. 13.7 - MICRO INQUIRY What would be the outcome if an...Ch. 13.7 - MICRO INQUIRY Why would it be impossible for...Ch. 13.7 - MICRO INQUIRY What provides the energy to fuel...Ch. 13.7 - Retrieve, Infer, Apply In which direction are...Ch. 13.7 - Retrieve, Infer, Apply Briefly describe the...Ch. 13.7 - Retrieve, Infer, Apply What are the translational...Ch. 13.7 - Retrieve, Infer, Apply Tabulate the nature and...Ch. 13.7 - Retrieve, Infer, Apply How many ATP and GTP...Ch. 13.8 - MICRO INQUIRY What are two distinguishing features...Ch. 13.8 - Retrieve, Infer, Apply What are molecular...Ch. 13.8 - Retrieve, Infer, Apply Would an intein-containing...Ch. 13.8 - Retrieve, Infer, Apply Give the major...Ch. 13.8 - Retrieve, Infer, Apply Which translocation or...Ch. 13.8 - Prob. 5RIACh. 13 - Streptomyces coelicolor has a linear chromosome....Ch. 13 - You have isolated several E. coli mutants: Mutant...Ch. 13 - DNA polymerase I (Pol I) of E. coli consists of...Ch. 13 - Prob. 4CHI
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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
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- 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
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