EBK GENETICS: FROM GENES TO GENOMES
EBK GENETICS: FROM GENES TO GENOMES
6th Edition
ISBN: 9781260041255
Author: HARTWELL
Publisher: MCGRAW HILL BOOK COMPANY
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Chapter 11, Problem 13P

You want to make a recombinant DNA in which a PCR product amplified from the human genome is inserted into a plasmid vector. The polylinker of this vector includes recognition sites for the enzymes EcoRI (5′ G^AATTC 3′) and BamHI (5′ G^GATCC 3′). (The ^ symbolizes the cut site in the DNA.) PCR primers that could amplify the fragment of human DNA are: 5′ GCTACTTCGCGTATTCCA 3′ and 5′CCCAAGTCCTAGCCGATA 3′.

a. Describe in detail how these primers would need to be modified to create a fragment of the human genome flanked by EcoRI sticky ends so that this fragment could be cloned easily into the plasmid vector. You will need to consider the fact that most restriction enzymes, including EcoRI, cannot cut DNA if the restriction site is directly at the end of the DNA molecule; the restriction enzyme recognition site must be at least six base pairs distant from the end.
b. Describe a potential feature of the PCR-amplified region of the human genome that could prevent you from using the strategy you described in part (a).
c. Now describe how the primers must be modified to create a human DNA fragment with an EcoRIcompatible single-stranded overhang at one end and a BamHI-compatible overhang at the other end. (Two possibilities exist; you need to describe only one. Assume that a BamHI site also must be at least six base pairs from the end of the DNA.) Why might you want to make such a fragment?
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Chapter 11 Solutions

EBK GENETICS: FROM GENES TO GENOMES

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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