You desperately need to join two DNA fragments, one with an EcoRl site and the other with a BamHI site. Both enzymes leave 5' overhangs cutting after the first base. EcoR/ cuts at gaattc, BamHI at ggatcc. Which of the following statements is true? A. You can not ligate these fragments. B. You can remove the overhangs with another restriction and ligate the blunted ends. C. You can fill in the ends with DNA polymerase and ligate the blunt ends. D. This can be done directly as the 5' overhangs anneal. E. You are better to replace EcoR/ with a related enzyme that leaves a ligatable 3' overhang.

You desperately need to join two DNA fragments, one with an EcoRl site and the other with a BamHI site. Both enzymes leave 5' overhangs cutting after the first base. EcoR/ cuts at gaattc, BamHI at ggatcc. Which of the following statements is true? A. You can not ligate these fragments. B. You can remove the overhangs with another restriction and ligate the blunted ends. C. You can fill in the ends with DNA polymerase and ligate the blunt ends. D. This can be done directly as the 5' overhangs anneal. E. You are better to replace EcoR/ with a related enzyme that leaves a ligatable 3' overhang.

Human Heredity: Principles and Issues (MindTap Course List)

11th Edition

ISBN:9781305251052

Author:Michael Cummings

Publisher:Michael Cummings

Chapter13: An Introduction To Genetic Technology

Section: Chapter Questions

Problem 8QP

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You are trying to clone a gene, You have successfully isolated it from the genomic DNA of an organism using the Hindill restriction enzyme. You then take a plasmid with a single EcoRI restriction site and cleave it with EcoRI. You combine these two fragments and treat them with DNA ligase. Answer the two questions below. a. Does the cloning reaction succeed as described? If so, what is the product obtained? b. Explain your answer above,
A. A plasmid is shown with the locations of various restriction enzyme sites labeled. If you cut the plasmid with Xhol and Xbal, which lane of the agarose gel represents the DNA fragments you would expect from the digestion? B. If you now decide to cut the plasmid with EcoRI, how many fragments will be produced and what will their sizes be? C. When running DNA samples on agarose gel, an electric field is applied. Towards which electrode will the DNA migrate and why?
A 12 kb linear DNA fragment is subject to single or double RE digest and agarose gelelectrophoresis, to yield the gel profile shown below. The first lane contains the size marker(M).a) Explain how the name of the enzyme EcoRI is derived.b) How many sites are there for EcoRI and PvuII respectively on this DNA fragment?c) Use the sizes of the DNA bands on the gel to compile a restriction enzyme map of the DNAfragment. Indicate the positions of the restriction enzymes sites for EcoRI and PvuII on themap.
When joining two or more DNA fragments, a researcher can adjust the sequence at the junction in a variety of subtle ways, as seen in the following exercises.(a) Draw the structure of each end of a linear DNA fragment produced by an EcoRI restriction digest (include those sequences remaining from the EcoRI recognition sequence).(b) Draw the structure resulting from the reaction of this end sequence with DNA polymerase I and the four deoxynucleoside triphosphates.(c) Draw the sequence produced at the junction that arises if two ends with the structure derived in (b) are ligated (d) Draw the structure produced if the structure derived in (a) is treated with a nuclease that degrades only single-stranded DNA.(e) Draw the sequence of the junction produced if an end with structure (b) is ligated to an end with structure (d).(f) Draw the structure of the end of a linear DNA fragment that was produced by a PvuII restriction digest (include those sequences remaining from the PvuII recognition…
You digested genomic dna with two restriction enzymes, Hinf I and Rsa I. These enzymes are used because: a. These enzymes recognize sequences TTAGGG present in TRF (terminal restriction fragment) at the ends of human chromosomes. b. These enzymes do not recognize sequences present in human telomeres' TRF (terminal restriction fragment). c. These enzymes are not expensive and very stable, making it possible to use them in a student laboratory course. d. These enzymes will not cut human genomic DNA
Consider a partial restriction digestion, in which genomic DNA is exposed to a small, limiting amount ofa restriction enzyme for a very short period of time.a. Would the resultant fragments be longer or shorteror the same size as those produced by a completedigestion?b. If you prepared genomic DNA from a tissue sample containing millions of cells, would the fragments produced by partial digestion of DNA fromall of these cells be the same or different?
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) Is the original molecule linear or circular?(b) Draw a map of restriction sites (showing distances between sites) that isconsistent with the data given.(c) How many additional maps are compatible with the data?(d) What would have to be done to locate the cleavage sites unambiguouslywith respect to each other?
How would the results of this activity have been different if the DNA sequence you digested were circular instead of linear? When restriction digests are performed and analyzed in the laboratory, one lane of the agarose gel is loaded with DNA ladders, which are DNA fragments of known sizes. What is the point of including these size standards?
Refer to the succeeding diagram of two DNA samples (A, B) with the specific restriction enzyme(RE) recognition sites marked with arrows. A spontaneous mutation (insertion) occurred in DNAsample A and introduced an additional restriction site. 1. Design two (2) DNA probes to be used in RFLP analysis involving these two DNA samples.Draw/illustrate the region(s) in the DNA molecule which you will use as reference(homologous) sequences in the design of the probes, THAT -a. Will reveal RFLP polymorphism between the DNA samples after probedetection/visualization; andb. Will not reveal RFLP polymorphism between the DNA samples
When circular DNA is sequenced, the nucleotide base pairs are numbered starting from a fixed position on the DNA, all the way around, usually in a clockwise manner. a DNA molecule that is 3133 base pairs long is digested with RsaI restriction enzyme recognition sites at base numbers 366, 1534, and 2207. What are the sizes of the DNA fragments that will be produced after the DNA is digested with RsaI?
The figure below shows the recognition sequences and cleavage positions of three restriction enzymes.You plan to ligate DNA from two different sources. The target DNA is digested with BamHI,and the insert DNA is digested with BglII, and the resulting fragments mixed and incubatedwith DNA ligase. a) Write out the sequence (in double-stranded format) of the longest insert fragment that will result after BglII digestion, ensure the nature of the overhangs is clear.b) Write out the sequence (in double-stranded format) of the ligation product, with the insert fragment joined into the BamHI site of the target DNA. Use black for target sequences, and blue for insert sequences. c) Assume the ligation reaction was successful and you have generated a recombinant DNAmolecule. Which of the three enzymes listed above can be used to excise the insert DNAfrom the target? Motivate your answer.
in Cohen-Boyer's recombinant DNA procedure _______ must be used for both the bacterial DNA and the amphibian DNA _______. a. the same restrictions enzyme, so that the the restriction site are identical in the DNA of each species b. different restriction enzymes, so that the genes outside the restriction site are maintained c. the same restriction enzymes, to ensure that the newly formed DNA can replicate d. different restriction enzymes, to ensure that the newly introducted genes are maintained in the bacterial DNA