Suppose that your bacterial cell contained a plasmid. When you isolated "genomic" DNA, would you expect...
Suppose that your bacterial cell contained a plasmid. When you isolated "genomic" DNA, would you expect to obtain the plasmid DNA along with the chromosomal DNA? Explain.
Homework Answers
- Bacterial plasmids are generally independent, self-replicating, autonomous, and extra-chromosomal DNA molecules that exist along with the bacterial genome.
- The plasmids replicate on their own during bacterial genome replication or they might insert themselves into the bacterial DNA and replicate along with it. The plasmids that occur independently are called as "relaxed" plasmids whereas the latter is called "stringent" plasmid.
- So, in the case of bacteria containing stringent plasmid, one may also isolate the plasmid DNA embedded in the bacterial genome whereas, relaxed plasmids can be obtained separately.
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Suppose that your bacterial cell contained a plasmid. When you
isolated "genomic" DNA, would you expect...
The bacterial replication machinery has been isolated and examined in vitro. The best way to learn...
The bacterial replication machinery has been isolated and examined in vitro. The best way to learn about the replication apparatus is to isolate it and them examine how each part works by itself and in concert with the other parts. Kornberg and coworkers worked very hard purifying all enzymes required for replication Initiation to try and study the process in detail. They then took the bacterial chromosome and digested it and plasmid DNA with restriction endonucleases (see below). They then...
8. Your lab assistant has tried to construct a genomic library of recombinant plasmids using only...
8. Your lab assistant has tried to construct a genomic library of recombinant plasmids using only the EcoRI restriction enzyme, genomic DNA, a plasmid and DNA ligase. The plasmid used is a typical cloning plasmid with a multiple cloning site (MCS) in the middle of a Lacz gene. After transforming E. coli with the library, a few white colonies are observed but a most of the colonies are blue. Explain what happened and what you would tell your assistant to...
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You want to digest the phage genomic DNA you just isolated with a restriction enzyme called HaeIII. You find the enzyme and a 10X stock of HaeIII enzyme buffer in the freezer. How much buffer will you add if the total volume of your reaction is 50 microliters?
7. You have performed a cloning experiment in which you digested genomic DNA from starfish cells...
7. You have performed a cloning experiment in which you digested genomic DNA from starfish cells with an enzyme and ligated the fragments into plasmid vectors digested with the same enzyme. The plasmid you are using contains a gene for ampicillin resistance and an MCS located within the Lacz gene. For the ligation, you added three times as much of the digested plasmid as the insert. After introducing the ligated DNA into cells, you observe 200 blue colonies and 33...
Can you answer these questions pleasee: If we performed electrophoretic separation (AGE) of human genomic DNA...
Can you answer these questions
pleasee: If we performed electrophoretic separation (AGE) of human
genomic DNA (buccal swab extraction), and we didn’t perform any
restriction digestion, answer the following:
a)In your opinion, which of the loaded samples has the highest
and lowest concentration of DNA, respectively?
b)Indicate the lanes in which DNA is degraded and explain your
answer.
c)Take a look into the lane “cut Plasmid”. Explain the appearance
of two bands. What kind of a technique was performed on...
You have cut genomic DNA with AvrII and wish to ligate it into pBluescript. However, there...
You have cut genomic DNA with AvrII and wish to ligate it into pBluescript. However, there is no AvrII site in the multiple cloning region of this plasmid. What other restriction enzyme could you use to cut the plasmid’s multiple cloning region that would produce compatible ends for ligation with your genomic DNA? (Hint: Look up the Compatible Cohesive Ends chart at New England Biolabs website and confer with pBluescript map) (1 point)
7. Explain the procedure for cloning DNA fragment into the plasmid PBR322 (shown on the right) (S...
7. Explain the procedure for cloning DNA fragment into the plasmid PBR322 (shown on the right) (S pts.). The gene fragment of interest was obtained by digestion of chromosomal DNA with the restriction enzyme Sall and subsequent purification using agarose gel electrophoresis. Which antiblotic would you use in the final step of the cloning procedure, and Pst why? EcoR Sal Ampicillin Tetracycline resistanica(Ter Amp) PBR322 4,361 bp) Origin of replicatiorn (ori Pvull 8. Assume that your gene fragment from question...
The exogenous DNA used in bacterial transformation can be, RONA mRNA molecule engineered plasmid red fluorescent...
The exogenous DNA used in bacterial transformation can be, RONA mRNA molecule engineered plasmid red fluorescent protein Incorrect Question 4 0/0.5 pts Bacteria that did not receive a plasmid are put on an LB plate that DOES contain ampicillin. What do you expect to happen? the bacteria will create a lawn the bacteria will not grow a few colonies will be seen http:/misac.instructure.com couro/87588/quizzes/163615 7/29/2020 Review Que: BIOL-8-05-12561.202010 Transformation efficiency. Concentration of plasmid DNA. Question 8 0.5/0.5 pts Super coiled...
You isolate plasmid DNA from bacteria (Questions 7-10) 7) A plasmid is an extrachromosomal circul...
10 please and 7
You isolate plasmid DNA from bacteria (Questions 7-10) 7) A plasmid is an extrachromosomal circular DNA frequently found in prokaryotes. Aside from being smaller, how is it different from the prokaryotic genome? You place equal amounts of plasmid DNA in 4 different tubes and incubate the DNA with increasing amounts of the enzyme topoisomerase I for 1 hour (0 enzyme units 0.25 enzyme units, 0.5 enzyme units and 1 enzyme unit). You then analyze the plasmid...
invas (2014) You prepare bacterial cell extracts by lysing the cells and removing insoluble debris via...
invas (2014) You prepare bacterial cell extracts by lysing the cells and removing insoluble debris via centrifugation. These extracts provide the proteins required for DNA replication. Your DNA template is a small, double-stranded circular piece of DNA (a plasmid) with a single origin of replication and a single replication termination site. The termination site is on the opposite side of the plasmid from the origin. origin plasmid termination Figure 6-11 What part of the DNA replication process would be most...
The bacterial replication machinery has been isolated and examined in vitro. The best way to learn about the replication apparatus is to isolate it and them examine how each part works by itself and in concert with the other parts. Kornberg and coworkers worked very hard purifying all enzymes required for replication Initiation to try and study the process in detail. They then took the bacterial chromosome and digested it and plasmid DNA with restriction endonucleases (see below). They then...
8. Your lab assistant has tried to construct a genomic library of recombinant plasmids using only the EcoRI restriction enzyme, genomic DNA, a plasmid and DNA ligase. The plasmid used is a typical cloning plasmid with a multiple cloning site (MCS) in the middle of a Lacz gene. After transforming E. coli with the library, a few white colonies are observed but a most of the colonies are blue. Explain what happened and what you would tell your assistant to...
7. You have performed a cloning experiment in which you digested genomic DNA from starfish cells with an enzyme and ligated the fragments into plasmid vectors digested with the same enzyme. The plasmid you are using contains a gene for ampicillin resistance and an MCS located within the Lacz gene. For the ligation, you added three times as much of the digested plasmid as the insert. After introducing the ligated DNA into cells, you observe 200 blue colonies and 33...
Can you answer these questions
pleasee: If we performed electrophoretic separation (AGE) of human
genomic DNA (buccal swab extraction), and we didn’t perform any
restriction digestion, answer the following:
a)In your opinion, which of the loaded samples has the highest
and lowest concentration of DNA, respectively?
b)Indicate the lanes in which DNA is degraded and explain your
answer.
c)Take a look into the lane “cut Plasmid”. Explain the appearance
of two bands. What kind of a technique was performed on...
7. Explain the procedure for cloning DNA fragment into the plasmid PBR322 (shown on the right) (S pts.). The gene fragment of interest was obtained by digestion of chromosomal DNA with the restriction enzyme Sall and subsequent purification using agarose gel electrophoresis. Which antiblotic would you use in the final step of the cloning procedure, and Pst why? EcoR Sal Ampicillin Tetracycline resistanica(Ter Amp) PBR322 4,361 bp) Origin of replicatiorn (ori Pvull 8. Assume that your gene fragment from question...
The exogenous DNA used in bacterial transformation can be, RONA mRNA molecule engineered plasmid red fluorescent protein Incorrect Question 4 0/0.5 pts Bacteria that did not receive a plasmid are put on an LB plate that DOES contain ampicillin. What do you expect to happen? the bacteria will create a lawn the bacteria will not grow a few colonies will be seen http:/misac.instructure.com couro/87588/quizzes/163615 7/29/2020 Review Que: BIOL-8-05-12561.202010 Transformation efficiency. Concentration of plasmid DNA. Question 8 0.5/0.5 pts Super coiled...
10 please and 7
You isolate plasmid DNA from bacteria (Questions 7-10) 7) A plasmid is an extrachromosomal circular DNA frequently found in prokaryotes. Aside from being smaller, how is it different from the prokaryotic genome? You place equal amounts of plasmid DNA in 4 different tubes and incubate the DNA with increasing amounts of the enzyme topoisomerase I for 1 hour (0 enzyme units 0.25 enzyme units, 0.5 enzyme units and 1 enzyme unit). You then analyze the plasmid...
invas (2014) You prepare bacterial cell extracts by lysing the cells and removing insoluble debris via centrifugation. These extracts provide the proteins required for DNA replication. Your DNA template is a small, double-stranded circular piece of DNA (a plasmid) with a single origin of replication and a single replication termination site. The termination site is on the opposite side of the plasmid from the origin. origin plasmid termination Figure 6-11 What part of the DNA replication process would be most...
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