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 11, Problem 24P
| a. | In Fig. 11.17b, PCR is performed to amplify genomic DNA for genotyping on microarrays. This PCR reaction requires only a single primer, but normally, PCR requires two primers. Why does only a single primer suffice in this case? |
| b. | Again in Fig. 11.17b, genomic DNA was cut with a restriction enzyme before its PCR amplification. What kind of restriction enzyme would be most effective for this purpose: Would it create sticky ends or blunt ends? Would it recognize a site made of 4 bp, 6 bp, or 8 bp? |
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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?
You are tasked with amplifying a gene of interest with PCR.
a. After performing a Southern blot, you see no signal on your gel. What could have
happened with your PCR?
b. After troubleshooting your PCR, you now want to perform Reverse Transcriptase
(RT) PCR to look at MRNA expression. Assuming the primers in 2a were not the
problem for your DNA and using the same primers as 2a, you see no signal on
your Southern blot. What are possible causes?
Explain why DNA ladders are usually included during gel electrophoresis.
One aspect of PCR that can be modified is the annealing temperature. In general, higher annealing temperatures show more specificity towards a single template, whereas lower annealing temperatures show less specificity and may bind to multiple regions throughout the genome. Discuss how using an annealing temperature that is too high or too low might influence the results of a PCR assay (and gel electrophoresis results) such as the one used in this study.
Chapter 11 Solutions
Genetics: From Genes to Genomes
Ch. 11 - Choose the phrase from the right column that best...Ch. 11 - Would you characterize the pattern of inheritance...Ch. 11 - Would you be more likely to find single nucleotide...Ch. 11 - A recent estimate of the rate of base...Ch. 11 - If you examine Fig. 11.5 closely, you will note...Ch. 11 - Approximately 50 million SNPs have thus far been...Ch. 11 - Mutations at simple sequence repeat SSR loci occur...Ch. 11 - Humans and gorillas last shared a common ancestor...Ch. 11 - In 2015, an international team of scientists...Ch. 11 - Using PCR, you want to amplify an approximately 1...
Ch. 11 - Prob. 11PCh. 11 - The previous problem raises several interesting...Ch. 11 - You want to make a recombinant DNA in which a PCR...Ch. 11 - You sequence a PCR product amplified from a...Ch. 11 - Prob. 15PCh. 11 - The trinucleotide repeat region of the Huntington...Ch. 11 - Sperm samples were taken from two men just...Ch. 11 - Prob. 18PCh. 11 - a. It is possible to perform DNA fingerprinting...Ch. 11 - On July 17, 1918, Tsar Nicholas II; his wife the...Ch. 11 - The figure that follows shows DNA fingerprint...Ch. 11 - Microarrays were used to determine the genotypes...Ch. 11 - A partial sequence of the wild-type HbA allele is...Ch. 11 - a. In Fig. 11.17b, PCR is performed to amplify...Ch. 11 - The following figure shows a partial microarray...Ch. 11 - Scientists were surprised to discover recently...Ch. 11 - The microarray shown in Problem 25 analyzes...Ch. 11 - The figure that follows shows the pedigree of a...Ch. 11 - One of the difficulties faced by human geneticists...Ch. 11 - Now consider a mating between consanguineous...Ch. 11 - The pedigree shown in Fig. 11.22 was crucial to...Ch. 11 - You have identified a SNP marker that in one large...Ch. 11 - The pedigrees indicated here were obtained with...Ch. 11 - Approximately 3 of the population carries a mutant...Ch. 11 - The drug ivacaftor has recently been developed to...Ch. 11 - In the high-throughput DNA sequencing protocol...Ch. 11 - A researcher sequences the whole exome of a...Ch. 11 - As explained in the text, the cause of many...Ch. 11 - Figure 11.26 portrayed the analysis of Miller...Ch. 11 - A research paper published in the summer of 2012...Ch. 11 - Table 11.2 and Fig. 11.27 together portray the...Ch. 11 - The human RefSeq of the entire first exon of a...Ch. 11 - Mutations in the HPRT1 gene in humans result in at...Ch. 11 - Prob. 44P
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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 Hindlll 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.(2 points Does the cloning reaction succeed as described? If so, what is the product obtained? b. Explain your answer above.arrow_forwardIrreversible organ failure remains a worldwide concern as demand for transplantable organs far outpaces the available supply. To overcome this shortage, xenotransplantation, where tissues or organs from animals are transplanted into humans has been tested with limited success since the early 1900's. Factors affecting the outcome of transplantation across the xenogeneic barrier include both humoral (innate) and cell-mediated (innate/adaptive) immune responses. Despite the development of strategies to achieve immunological tolerance, xenotransplantation is not yet a clinical reality. Indeed, while organ rejection can be avoided, the risk of an infectious or zoonotic disease spreading from animals to humans raises further concerns about the clinical application of xenotransplantation. By the late 1990s, the advent of stem cell research had sparked a new direction in the field of regenerative medicine. As an alternative to generating organs from induced pluripotent stem cells (iPSCS) in…arrow_forwardOn the gel shown below are four DNA samples. Samples A to C are taken from tissues of landslide victims that are being identified, while sample D came from a hair sample brought by a mother looking for the remains of her son. (see img) i. If similar band patterns in a gel are created using the same restriction enzyme, what does that tell you about the DNA sequence of the samples? ii. In sample C, only two fragments were created. How many restriction sites (regions where enzymes cut) are present in sample C?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_forwarda. What determines the size (length) of the primary PCR product? b. What might a successful gel check of a PCR reaction look like?arrow_forwardWhat is the principle of the Restriction Fragment Length Polymorphism (RFLP) in the diagnosis of human diseases? O a. PCR product of a gene is different from the expected one b. The size of a recombinant DNA is different from the expected one OC. The size of a band digested by specific restriction enzymes is different from the expected one O d. The DNA band detected by Southern blot is different from that by Northern blotarrow_forward
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- The same restriction endonuclease must be used to excise the foreign DNA and bacterial DNA. Select one: True False Why? Select one: a. It must use different restriction endonucleases because the bacterial and foreign DNA sequences are different. b. It must use same restriction endonuclease so that the restriction sites are identical in both foreign and bacterial DNA. c. It must use same restriction endonuclease so that the restriction sites are different in both foreign and bacterial DNA. d. It must use different restriction endonucleases because the bacterial and foreign DNA sequences are exactly the same.arrow_forwardMost PCR reactions do not use the more expensive types of DNA polymerase, which have DNA proofreading. How might this be a problem in accurately copying specific DNA sequences of the target gene?arrow_forwarda. What type of nucleic acid and from what species would the scientist use to begin construction of her genomic DNA library? b. From what tissue would she isolate this nucleic acid? c. What type of reagent would the scientist use to cut the genome into appropriately sized fragments? d. What size nucleic acid fragments would one aim to prepare for the library construction so as to to avoid having to screen an overwhelming number of clones? e. Into what vector would the scientist ligate her genomic DNA fragments? f. What organism would the scientist use to propagate the clones of her genomic DNA library? g. From the information given in the problem determine what probe could be used to screen the scientist's library to find her clone of interest ?arrow_forward
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