Genetics: From Genes to Genomes
6th Edition
ISBN: 9781259700903
Author: Leland Hartwell Dr., Michael L. Goldberg Professor Dr., Janice Fischer, Leroy Hood Dr.
Publisher: McGraw-Hill Education
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Chapter 9, Problem 23P
| a. | To make a genomic library useful for sequencing an entire genome, why would you ordinarily fragment the genomic DNA by mechanical shearing forces like sonication rather than by cutting the DNA with a restriction enzyme? |
| b. | Suppose that you wanted to make a genomic library to determine the complete sequence of a newly discovered organism’s genome, but you did not have a sonicator readily available. Explain how you could nonetheless use two or more restriction enzymes to make libraries whose clones could be sequenced so that a computer could assemble the genomic sequence. |
| c. | Suppose you only had a single restriction enzyme available, and you want to make a single genomic library from which you could assemble the genomic sequence. How might you be able to achieve this goal? (Hint: See Problem 9.) To make this library, would it be preferable to use a restriction enzyme that recognizes a 4-base, 6-base, or 8-base sequence of DNA? |
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a To make a genomic library useful for sequencingan entire genome, why would you ordinarily fragment the genomic DNA by mechanical shearingforces like sonication rather than by cutting theDNA with a restriction enzyme?b. Suppose that you wanted to make a genomic library to determine the complete sequence of anewly discovered organism’s genome, but you didnot have a sonicator readily available. Explain howyou could nonetheless use two or more restrictionenzymes to make libraries whose clones could besequenced so that a computer could assemble thegenomic sequence.c. Suppose you only had a single restriction enzymeavailable, and you want to make a single genomiclibrary from which you could assemble the genomic sequence. How might you be able to achievethis goal? (Hint: See Problem 9.) To make this library, would it be preferable to use a restriction enzyme that recognizes a 4-base, 6-base, or 8-basesequence of DNA?
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?
Suppose that a human genomic library is prepared by exhaustive digestion of human DNA with the EcoRI restriction enzyme. Fragments averaging about 4 kb in length would be generated. Is this procedure suitable for cloning large genes? Why or why not?
Chapter 9 Solutions
Genetics: From Genes to Genomes
Ch. 9 - Match each of the terms in the left column to the...Ch. 9 - For each of the restriction enzymes listed below:...Ch. 9 - The calculations of the average restriction...Ch. 9 - The DNA molecule whose entire sequence follows is...Ch. 9 - Why do longer DNA molecules move more slowly than...Ch. 9 - Agarose gels with different average pore sizes are...Ch. 9 - The following picture shows the ethidium...Ch. 9 - The linear bacteriophage genomic DNA has at each...Ch. 9 - Consider a partial restriction digestion, in which...Ch. 9 - The text stated that molecular biologists have...
Ch. 9 - a. What is the purpose of molecular cloning? b....Ch. 9 - a. DNA polymerase b. RNA polymerase c. A...Ch. 9 - Is it possible that two different restriction...Ch. 9 - A plasmid vector pBS281 is cleaved by the enzyme...Ch. 9 - A recombinant DNA molecule is constructed using a...Ch. 9 - Suppose you are using a plasmid cloning vector...Ch. 9 - Prob. 17PCh. 9 - The lacZ gene from E. coli encodes the enzyme...Ch. 9 - Your undergraduate research advisor has assigned...Ch. 9 - Which of the enzymes from the following list would...Ch. 9 - You use the primer 5 GCCTCGAATCGGGTACC 3 to...Ch. 9 - a. To make a genomic library useful for sequencing...Ch. 9 - Problem 15 showed part of the sequence of the...Ch. 9 - Eukaryotic genomes are replete with repetitive...
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- How many fragments would you expect to be formed from digestion of a 2500 base pair long linear piece of DNA using a restriction enzyme with a 5 base pair recognition sequence?”arrow_forwardIf the GAATTC palindrome repeats are randomly found along the DNA strand, then what can you say about the sizes of the fragments that will be produced when the DNA is digested with a restriction enzyme that recognizes that sequence? How does the total length of the fragments relate to the size of the original DNA fragment?arrow_forwardWhen 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…arrow_forward
- 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,arrow_forwardFrom your knowledge about DNA microarray, answer the following: A- How DNA microarray is created? and why it is referred to as “hybridization technology”? B- Why RT-PCR is important in the sample preparation to perform expression microarray experiment? C- Mention the name and the color of the dyes used in expression microarray? D- If the expression microarray experiment was done with a normal sample and a suspected sample, after reading the color pattern resulted from the experiment it was recorded that “gene A22” is expressed in the suspected sample. The gene A22 is clinically linked to colon cancer. Answer the following: What is the expected color of the spot on the microarray which represents this gene? What is your interpretation of the suspected sample; is it a cancer sample or not and explain why?arrow_forwardIs it possible that two different restriction enzymes couldcut the human genome into exactly the same number offragments and with exactly the same distribution of fragment sizes, yet the ends produced by the two enzymescould not be joined together by DNA ligase? Explainarrow_forward
- If a pair of 10-mer primers are used in a PCR, what would be the number of expected binding sites in a human genomic DNA? (You can use approximate values.)arrow_forwardConsider 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?arrow_forwardA 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 bparrow_forward
- 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?arrow_forwardDuring PCR amplification in preparation for DNA sequencing, why were there different colors at the 3’ ends of the fragments produced? (What did these four colors represent?)arrow_forwardWhat is the main reason for using a cDNA library rather than a genomic library to isolate a human gene from which you wish to make large quantities of the human protein in bacteria? How will you identify clones of interest?arrow_forward
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