A piece of DNA 5.0 kb long is cloned and then cut out of the vector for analysis. This linearpiece of DNA is digested with two restriction enzymes, EcoRI and BamHI, individually and incombination, and the resulting fragment sizes are determined by electrophoresis. The results areas follows:Restriction fragment size4.5 kb; 0.5 kb3.0 kb; 2.0 kb2.5 kb; 2.0 kb; 0.5 kbEnzyme nameEcoRIВатHIEcoRI + BamHIConstruct a potential restriction map based on these results.

Restriction enzyme is any enzyme that cleaves DNA at a particular sequence. Example - BamHI, EcoRI,…

Answered: A piece of DNA 5.0 kb long is cloned…

Biology Today and Tomorrow without Physiology (MindTap Course List)

5th Edition

ISBN:9781305117396

Author:Cecie Starr, Christine Evers, Lisa Starr

Publisher:Cecie Starr, Christine Evers, Lisa Starr

Chapter10: Biotechnology

Section: Chapter Questions

Problem 10SQ

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A small DNA molecule was cleaved with several different restriction nucleases, and the size of each fragment was determined by gel electrophoresis. The following data were obtained.
A 22-kb piece of DNA has the following restriction sites:A batch of this DNA is first fully digested by HpaI alone, then another batch is fully digested by HindIII alone, and finally, a third batch is fully digested by both HpaI and HindIII together. The fragments resulting from each of the three digestions are placed in separate wells of an agarose gel, separated by gel electrophoresis, and stained by ethidium bromide. Draw the bands as they would appear on the gel.
The gene you are asked to clone is 30,000 bps in length. When you are choosing a suitable Restriction Endonuclease, what criteria about the enzyme can you deduce from the gene length?
All of the following are performed during restriction fragment length polymorphism analysis. 1. splitting of double-stranded into single-stranded DNA 2. gel electrophoresis 3. autoradiography 4. immersion in radioactive probes 5. digestion of DNA with restriction endonucleases 6. use of a positive charge to transfer single-stranded DNA from a gel to a membrane. The correct sequence of these operations is what
Which vectors can be used to clone a continuous fragment of DNA with the following lengths? a. 4 kb b. 20 kb c. 35 kb d. 100 kb
Above are the results of gel electrophoresis following digestion with restriction enzymes. What is the total length of the DNA fragment? Which enzyme, HindIII or EcoRI, produced a larger fragment? What part of DNA causes it to be negatively charged in order for electrophoresis to work?
If you were to isolate genomic DNA from two people and perform restriction enzyme digestion followed by agarose gel electrophoresis, would this be an effective way to distinguish one person from the other? Explain. If not, what is another technique you might use to distinguish the two people?
A purified piece of DNA of 1650bp is cut with Hind III and Pst I separately and then with both enzymes together. The following are the observations in gel electrophoresis (fragments sizes are in bp). HindIII 300 500 850 PstI 100 600 950 HindIII and PstI 100 250 300 400 600 Use this information and make a restriction map of DNA.
A 2.0kb bacterial plasmid ‘BS1030’ is digested with the restriction endonuclease Sau3A; the plasmid map is depicted in the diagram below and the Sau3A (S) restriction sites are indicated. Which of the following DNA fragments do you expect to see on an agarose gel when you run Sau3A-digested plasmid ‘BS1030’ DNA? a. 250 bp, 450 bp, 550 bp, 1.1 kb, 1.5 kb and 2.0 kb b. 2.0kb c. 250 bp, 400 bp, 450 bp, 500 bp and 550 bp d. 100 bp, 200 bp, 250 bp, 400 bp, 500 bp and 550 bp
When the restriction endonuclease EcoRI is used to digest a 10 kb DNA fragment, it produces 4 kb and 6 kb-sized fragments. Digesting the 10 kb fragment with BamHI yields three fragments, each ranging in size from one to three and a half kilobytes. Four pieces of 0.5, 1, 3 and 5.5 kb are formed after using both enzymes. Create a restriction map for this 10 kb piece of DNA using the information you have collected. Make a note of where the two enzymes cut, as well as the distances between the enzymes.
In relation to the use of restriction enzymes in recombinant DNA technology, answer the following: You have accidentally torn the labels off two tubes (tube A and tube B), each containing a different plasmid, now you do not know which plasmid is in which tube. Fortunately, you have restriction maps for both plasmids, shown in Figure below. You have the opportunity to test just one sample from one of your tubes. By utilizing agarose gel electrophoresis technique, which restriction enzyme OR combination of restriction enzymes would you use in this experiment to determine which plasmid is found in which tube?. (Hint: if you use Hind III restriction enzyme you are going to get ONE single fragment with a molecular size of → 0.5+0.3+0.2+0.4+1+1 = 3.4 kb).
You are performing an experiment using CRISPR-cas9 to genetically modify the LacZ gene of a culture of E. coli. After you run the experiment, you decide to use gel electrophoresis to genotype the different bacterial cultures to determine if the gene editing was successful. How could your electrophoresis results confirm that the PCR was successful? And how could your electrophoresis results confirm that you successfully extracted genomic DNA from your bacterial samples?

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