Question 1

Compare and contrast genomic libraries with cDNA libraries.

Accepted Answer

The answer of Compare and contrast genomic libraries with cDNA...

Question 2

Describe the principle of ion-exchange chromatography. Describe the behavior of a positively charged protein and a negatively charged protein in a column containing positively charged resin during sample loading and elution.

Accepted Answer

The answer of Describe the principle of ion-exchange chromatography. Describe...

Question 3

Describe the principle of gel filtration chromatography. For two proteins of different size, illustrate in which fraction they will elute.

Accepted Answer

The answer of Describe the principle of gel filtration chromatography....

Question 4

Describe the principle of antibody-affinity chromatography. For one protein that is recognized by the antibody and a second protein that is not, illustrate at which point each protein will be released from the column.

Accepted Answer

The answer of Describe the principle of antibody-affinity chromatography. For...

Question 5

Diagram the process for creating a double-stranded cDNA.

Accepted Answer

The answer of Diagram the process for creating a double-stranded...

Question 6

Explain why DNA polymerase from a thermophilic bacterium is used in PCR instead of DNA polymerase I from E. coli.

Accepted Answer

The answer of Explain why DNA polymerase from a thermophilic...

Question 7

Compare and contrast the principles of autoradiography and phosphorimaging.

Accepted Answer

The answer of Compare and contrast the principles of autoradiography...

Question 8

What are radioisotopes and why are they useful in molecular biology research?

Accepted Answer

The answer of What are radioisotopes and why are they...

Question 9

Define the term "probe" and describe two important characteristics of a probe.

Accepted Answer

The answer of Define the term "probe" and describe two...

Question 10

What are the relative advantages and disadvantages of end-labeling versus internal-labeling a probe?

Accepted Answer

The answer of What are the relative advantages and disadvantages...

Question 11

(a) Outline the steps that would be necessary to generate a cDNA library from all of the genes expressed by adenocarcinoma cells of a breast tumor. (b) How could tumor-specific transcripts be identified from this library? (Hint: think about how you could use non-cancerous breast epithelial cells as a comparison.)

Accepted Answer

The answer of (a) Outline the steps that would be...

Question 12

Compare and contrast degenerate probes and EST-based probes.

Accepted Answer

The answer of Compare and contrast degenerate probes and EST-based...

Question 13

Diagram a strategy for screening a cDNA library.

Accepted Answer

The answer of Diagram a strategy for screening a cDNA...

Question 14

If you had an antibody to the protein encoded by an unknown gene of interest, what type of library could you screen to identify the sequence of the gene? Using a diagram, explain how this would work.

Accepted Answer

The answer of If you had an antibody to the...

Question 15

Show how to use restriction mapping to determine the orientation of a restriction fragment ligated into a restriction site in a vector.

Accepted Answer

The answer of Show how to use restriction mapping to...

Question 16

Diagram the process of Southern blotting and probing to detect a DNA of interest. What kinds of information can be obtained from a Southern blot?

Accepted Answer

The answer of Diagram the process of Southern blotting and...

Question 17

Compare and contrast the key characteristics of automated sequencing using the Sanger method and 454 pyrosequencing.

Accepted Answer

The answer of Compare and contrast the key characteristics of...

Question 18

You have attempted to ligate a 1.5 kb fragment of foreign DNA into the EcoRI site in the multiple cloning site of the 4.0 kb plasmid vector shown below:
‪  (a) After ligation you use the DNA in the ligation mixture to transform host bacteria. Why is it important to use host bacteria that are deficient for restriction-modification?
(b) You screen the bacteria that supposedly have been transformed with recombinant plasmid DNA. Some of the bacterial colonies growing on the nutrient agar plate that contains ampicillin and X-gal are white and some are blue. Explain these results.
(c) To confirm the presence of the foreign DNA insert, you perform EcoRI restriction endonuclease digests on DNA extracted from bacterial colonies. On the diagram of an agarose gel shown below, draw the pattern of bands (label their size in kb) you would expect to see for EcoRI-digested recombinant plasmid and EcoRI-digested non-recombinant plasmid vector, after electrophoresis and staining of the gel with ethidium bromide.

Accepted Answer

The answer of You have attempted to ligate a 1.5...

Question 19

A chromotography column in which oligo-dT is linked to an inert substance is useful in separating eukaryotic mRNA from other RNA molecules. On what principle does this column operate?

Accepted Answer

The answer of A chromotography column in which oligo-dT is...

Question 20

Starting with the nucleotide sequence of the human DNA ligase I gene, describe how you would search for a homologous gene in another organism whose genome has been sequenced, such as the pufferfish Tetraodon nigroviridis. Then, describe how you would obtain the protein and test it for ligase activity.

Accepted Answer

The answer of Starting with the nucleotide sequence of the...

Quiz 8: Recombinant Dna Technology and Molecular Cloning