Chapter 20: Problem 14
What advantages do cDNA libraries provide over genomic DNA libraries? Describe cloning applications where the use of a genomic library is necessary to provide information that a cDNA library cannot.
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Chapter 20: Problem 14
What advantages do cDNA libraries provide over genomic DNA libraries? Describe cloning applications where the use of a genomic library is necessary to provide information that a cDNA library cannot.
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The human insulin gene contains a number of sequences that are removed in the processing of the mRNA transcript. In spite of the fact that bacterial cells cannot excise these sequences from mRNA transcripts, explain how a gene like this can be cloned into a bacterial cell and produce insulin.
List the advantages and disadvantages of using plasmids as cloning vectors. What advantages do BACs and YACs provide over plasmids as cloning vectors?
What roles do restriction enzymes, vectors, and host cells play in recombinant DNA studies? What role does DNA ligase perform in a DNA cloning experiment? How does the action of DNA ligase differ from the function of restriction enzymes?
A human cDNA clone was isolated from a cDNA library using a homologous mouse gene as a probe. The clone was grown in \(E\) coll, miniprepped to isolate the \(c D N A\), and then amplificd by \(\mathrm{PCR}\) using primers designed to the sequence of the mouse gene. When the PCR reaction was run on an agarose gel, there were multiple weak bands present instead of one clear band. What are the possible reasons for this result? How can the experiment be improved so that one clear band is amplified next time?
Although many cloning applications involve introducing recombinant DNA into bacterial host cells, many other cell types are also used as hosts for recombinant DNA. Why?
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