Question 1

You want to isolate an insert from a vector using PvuII restriction sites as shown. What is the easiest way to isolate the insert away from the rest of the vector?

Accepted Answer

Digest the vector with a restriction endonuclease and make a library from the pieces.

Question 2

Which technique is best for creating a specific mutation in a piece of DNA?

Accepted Answer

Use a primer with a mismatched nucleotide.

Question 3

A DNA library is plated on a petri dish, transferred to a membrane, and washed with a fluorescent probe. This procedure is called ________.

Accepted Answer

Screening the library

Question 4

You have cloned the green fluorescent protein (GFP) gene next to the promoter for the proP gene. For what purpose have you done this?

Accepted Answer

To use GFP as a reporter to test the proP gene promoter activity

Question 5

What is the correct order for the three steps in the polymerase chain reaction (PCR), starting with a double stranded DNA sequence?

Accepted Answer

Denature the DNA, anneal primers, elongate the DNA

Question 6

Which of the following is NOT used when performing a Southern Blot?

Question 7

A patient is infected with Staphylococcus aureus. DNA is extracted, denatured, and PCR is performed with primers specific for a S. aureus gene. The denaturing, priming, and polymerase steps are repeated 40 times. The product of the reaction is run on an electrophoretic gel, but no bands are visible. Why not?

Accepted Answer

The patient is not infected with Staphylococcus aureus.

Question 8

The lacZ gene, whose colored product is easily seen, is inserted without its promoter into the middle of the coding region of the proP gene. The lacZ gene in this experiment is being used as a ________.

Accepted Answer

Reporter gene

Question 9

Which of the following is NOT a use for an oligonucleotide probe?

Accepted Answer

Performing site-directed mutagenesis on a particular gene

Question 10

You want to mutate an amino acid with a positively-charged R-group in the active site of enzyme X to one with a negatively-charged R-group. Which technique would be most effective?

Accepted Answer

Use a probe with a sequence mismatch that will replace the codon for the gene for enzyme X

Question 11

Which of the following sequences is considered a palindrome?

Accepted Answer

$5'\ CCCAAA\ 3'$

Question 12

Which step in the PCR process involves the separation of DNA strands?

Accepted Answer

DNA is heated to 95$^\circ$C

Question 13

A probe can be used in all of the following procedures EXCEPT . . .

Accepted Answer

Amplifying the number of copies of a selected DNA fragment

Question 14

You want to design an experiment to test the hypothesis that the glutamic acid (Glu) at a particular place in the sequence of the protein AmhE is important for bond formation. Which experiment would be most likely to give you that data you need?

Accepted Answer

Use a primer containing the sequence TTT rather than CTT to mutate the gene.

Question 15

Which of the following is the best way to determine whether a particular B. anthracis cell is pathogenic?

Accepted Answer

Use of reporter gene inserted somewhere on the plasmid

Question 16

Which of the following is a multi-step process that can be used to clone a specific gene?

Accepted Answer

Isolate a product under the gene you want to clone, screen clones for presence of a particular gene, transform host bacterial cells with cloning vector, digest electrophoresis of DNA fragments, ligate pieces of DNA into vector, digest organism's DNA and cloning vector

Question 17

Which of the following is NOT true about recombinant DNA formed with restriction enzymes NheI and StyI?

Accepted Answer

The DNA ligase would not recognize these sequences.

Question 18

Which of the following is the best description of a reporter gene?

Accepted Answer

A gene with a visible product used to report the activity of another gene's promoter

Question 19

Which of the following steps in the PCR (polymerase chain reaction) is in the correct order?

Accepted Answer

DNA is heated to 95$^\circ$C, primers are annealed to the 3' ends of the gene to be copied, DNA polymerase transcribes the DNA

Molecular Techniques in Microbiology: Cloning, Mutagenesis, and DNA Analysis

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