Question 1

Which of the following procedures can be used to avoid contamination with PCR reaction product (amplicon)when performing a PCR procedure?

Accepted Answer

A)  Use of positive displacement pipettes 
B)  Use of closed-tube methods 
C)  Physical separation of preamplification and postamplification rooms 
D)  All of the above 
A)  Use of positive displacement pipettes 
B)  Use of closed-tube methods 
C)  Physical separation of preamplification and postamplification rooms 
D)  All of the above D

Question 2

The increase in the quantifiable signal observed early on in real-time PCR is dependent upon the:

Accepted Answer

A)  amount of fluorescent dye added to the reaction. 
B)  initial amount of fluorescent primer added. 
C)  amount of fluorescent quenching. 
D)  initial amount of target DNA. 
A)  amount of fluorescent dye added to the reaction. 
B)  initial amount of fluorescent primer added. 
C)  amount of fluorescent quenching. 
D)  initial amount of target DNA. D

Question 3

You want to determine the accuracy of hybridization of a probe to a Southern blot to eliminate binding of the probe to incorrect target sequences.Which one of the following would be an appropriate positive control to assess assay sensitivity?

Accepted Answer

A)  Sequences complementary to the probe 
B)  Sequences that are not complementary to the probe 
C)  Water 
D)  Sequences from a housekeeping gene such as beta actin 
A)  Sequences complementary to the probe 
B)  Sequences that are not complementary to the probe 
C)  Water 
D)  Sequences from a housekeeping gene such as beta actin A

Question 4

Which one of the following is the name given to the probe types used in real-time PCR that change fluorescence through fluorescence resonance energy transfer (FRET)upon duplex formation?

Accepted Answer

A)  Hybridization 
B)  Hydrolysis 
C)  Mixed mechanism 
D)  Primer 
A)  Hybridization 
B)  Hydrolysis 
C)  Mixed mechanism 
D)  Primer

Question 5

Which one of the following statements regarding single-stranded conformational polymorphism (SSCP)analysis is incorrect?

Accepted Answer

A)  SSCP is used to assess a specimen for the presence of unknown variants in a nucleic acid sequence. 
B)  SSCP distinguishes altered gene sequences by detecting a change in the three-dimensional structure of a piece of single-stranded DNA. 
C)  SSCP is a polyacrylamide gel electrophoresis procedure. 
D)  In SSCP,separation of a PCR product is performed with a gel that includes a concentration gradient of denaturants. 
A)  SSCP is used to assess a specimen for the presence of unknown variants in a nucleic acid sequence. 
B)  SSCP distinguishes altered gene sequences by detecting a change in the three-dimensional structure of a piece of single-stranded DNA. 
C)  SSCP is a polyacrylamide gel electrophoresis procedure. 
D)  In SSCP,separation of a PCR product is performed with a gel that includes a concentration gradient of denaturants.

Question 6

In real-time PCR:

Accepted Answer

A)  an isolated mRNA and Thermus thermophilus is used. 
B)  one can observe amplified nucleic acid fragments as they are synthesized. 
C)  RNA:DNA hybrids are bound to a solid phase. 
D)  RNA targets are used in an isothermal reaction. 
A)  an isolated mRNA and Thermus thermophilus is used. 
B)  one can observe amplified nucleic acid fragments as they are synthesized. 
C)  RNA:DNA hybrids are bound to a solid phase. 
D)  RNA targets are used in an isothermal reaction.

Question 7

Which one of the following statements regarding DNA gel electrophoresis is true?

Accepted Answer

A)  It is the charge of the pyrimidines in the DNA molecule that gives DNA its net negative charge and ability to migrate on a gel. 
B)  Agarose gel can resolve 1 bp differences in DNA fragments and is best to use for DNA sequencing. 
C)  Polyacrylamide gel can separate smaller fragments of DNA with high resolution and is best for single-stranded nucleic acid separation. 
D)  Because of the negative charge of DNA,it will migrate toward the negative electrode during electrophoresis. 
A)  It is the charge of the pyrimidines in the DNA molecule that gives DNA its net negative charge and ability to migrate on a gel. 
B)  Agarose gel can resolve 1 bp differences in DNA fragments and is best to use for DNA sequencing. 
C)  Polyacrylamide gel can separate smaller fragments of DNA with high resolution and is best for single-stranded nucleic acid separation. 
D)  Because of the negative charge of DNA,it will migrate toward the negative electrode during electrophoresis.

Question 8

An example of a signal amplification technique (as opposed to a target or probe amplification technique)would be:

Accepted Answer

A)  polymerase chain reaction. 
B)  branched-chain amplification. 
C)  ligase chain reaction. 
D)  real-time PCR. 
A)  polymerase chain reaction. 
B)  branched-chain amplification. 
C)  ligase chain reaction. 
D)  real-time PCR.

Question 9

What type of assay is hybrid capture?

Accepted Answer

A)  Solid-phase hybridization 
B)  Signal amplification 
C)  Transcription-mediated amplification 
D)  Solution-phase hybridization 
A)  Solid-phase hybridization 
B)  Signal amplification 
C)  Transcription-mediated amplification 
D)  Solution-phase hybridization

Question 10

In a pyrosequencing reaction,the incorporation of a nucleotide releases _____.Eventually,visible light is produced by an enzyme reaction.

Accepted Answer

A)  pyrophosphate 
B)  ATP 
C)  luciferase 
D)  a primer 
A)  pyrophosphate 
B)  ATP 
C)  luciferase 
D)  a primer

Question 11

If RNA is to be used in a PCR amplification procedure,what is the initial step that must be performed?

Accepted Answer

A)  The RNA must be denatured to form single strands for annealing to primers. 
B)  RNA should never be used in a PCR reaction because thermostable enzymes cannot synthesize strands to anneal to it. 
C)  A reverse transcription procedure must be performed to form cDNA. 
D)  RNA must first be treated with RNases to remove interfering substances from the target. 
A)  The RNA must be denatured to form single strands for annealing to primers. 
B)  RNA should never be used in a PCR reaction because thermostable enzymes cannot synthesize strands to anneal to it. 
C)  A reverse transcription procedure must be performed to form cDNA. 
D)  RNA must first be treated with RNases to remove interfering substances from the target.

Question 12

An enzyme that hydrolyzes one or more phosphodiester bonds in nucleic acid polymers is called a:

Accepted Answer

A)  polymerase. 
B)  nuclease. 
C)  ligase. 
D)  reverse transcriptase. 
A)  polymerase. 
B)  nuclease. 
C)  ligase. 
D)  reverse transcriptase.

Question 13

What is the key enzyme used in a polymerase chain reaction (PCR)?

Accepted Answer

A)  Restriction enzyme 
B)  Lysozyme 
C)  Polymerase 
D)  Transcriptase 
A)  Restriction enzyme 
B)  Lysozyme 
C)  Polymerase 
D)  Transcriptase

Question 14

Gene expression microarrays quantify:

Accepted Answer

A)  a single copy of a nucleic acid target. 
B)  relative amounts of different messenger RNAs in samples. 
C)  RNA promoter regions that are adjacent to an inserted DNA sequence. 
D)  the number of specific base pairs in a DNA target. 
A)  a single copy of a nucleic acid target. 
B)  relative amounts of different messenger RNAs in samples. 
C)  RNA promoter regions that are adjacent to an inserted DNA sequence. 
D)  the number of specific base pairs in a DNA target.

Question 15

The purpose of restriction fragment polymorphism analysis (RFLP)is to:

Accepted Answer

A)  determine the purity of DNA isolation. 
B)  evaluate the integrity of isolated genomic DNA. 
C)  assess the exonuclease activity of DNA polymerase. 
D)  identify the presence of mutations or sequence changes. 
A)  determine the purity of DNA isolation. 
B)  evaluate the integrity of isolated genomic DNA. 
C)  assess the exonuclease activity of DNA polymerase. 
D)  identify the presence of mutations or sequence changes.

Question 16

How does a dideoxynucleotide (ddNTP)that is used in DNA sequencing methods differ from a typical deoxynucleotide (dNTP)present in normal DNA?

Accepted Answer

A)  ddNTPs have a hydroxyl group at the 2' carbon 
B)  ddNTPs have a hydroxyl group at the 3' carbon 
C)  ddNTPs lack a hydroxyl group at the 3' carbon 
D)  ddNTPs lack a hydroxyl group at the 5' carbon 
A)  ddNTPs have a hydroxyl group at the 2' carbon 
B)  ddNTPs have a hydroxyl group at the 3' carbon 
C)  ddNTPs lack a hydroxyl group at the 3' carbon 
D)  ddNTPs lack a hydroxyl group at the 5' carbon

Question 17

False positive results in molecular testing are most likely caused by which one of the following?

Accepted Answer

A)  Inhibitors present in the sample 
B)  Contamination with amplicon 
C)  Lengthy specimen prep time 
D)  Lengthy patient prep time 
A)  Inhibitors present in the sample 
B)  Contamination with amplicon 
C)  Lengthy specimen prep time 
D)  Lengthy patient prep time

Question 18

In the dideoxy-termination sequencing method,what causes the termination of the newly synthesized DNA strand?

Accepted Answer

A)  Addition of a dideoxynucleotide 
B)  Deletion of a deoxyribonucleotide 
C)  Addition of a deoxyribonucleotide 
D)  Addition of a ribonucleotide 
A)  Addition of a dideoxynucleotide 
B)  Deletion of a deoxyribonucleotide 
C)  Addition of a deoxyribonucleotide 
D)  Addition of a ribonucleotide

Question 19

Regarding detection methods,the most common and preferred sequence-specific label for probes used in nucleic acid analysis is a(n)_____ label.

Accepted Answer

A)  enzyme 
B)  radioactive 
C)  fluorescent 
D)  colorimetric 
A)  enzyme 
B)  radioactive 
C)  fluorescent 
D)  colorimetric

Question 20

What is the purpose of the labeled oligonucleotide probe used in hybridization assays?

Accepted Answer

A)  To stabilize the assay 
B)  To cut the DNA into smaller-size fragments 
C)  To reduce the background of the assay 
D)  To detect the presence of a specific nucleotide sequence 
A)  To stabilize the assay 
B)  To cut the DNA into smaller-size fragments 
C)  To reduce the background of the assay 
D)  To detect the presence of a specific nucleotide sequence

Exam 48: Nucleic Acid Techniques and Applications

Which of the following procedures can be used to avoid contamination with PCR reaction product (amplicon)when performing a PCR procedure?

The increase in the quantifiable signal observed early on in real-time PCR is dependent upon the:

You want to determine the accuracy of hybridization of a probe to a Southern blot to eliminate binding of the probe to incorrect target sequences.Which one of the following would be an appropriate positive control to assess assay sensitivity?

Which one of the following is the name given to the probe types used in real-time PCR that change fluorescence through fluorescence resonance energy transfer (FRET)upon duplex formation?

Which one of the following statements regarding single-stranded conformational polymorphism (SSCP)analysis is incorrect?

In real-time PCR:

Which one of the following statements regarding DNA gel electrophoresis is true?

An example of a signal amplification technique (as opposed to a target or probe amplification technique)would be:

What type of assay is hybrid capture?

In a pyrosequencing reaction,the incorporation of a nucleotide releases _____.Eventually,visible light is produced by an enzyme reaction.

If RNA is to be used in a PCR amplification procedure,what is the initial step that must be performed?

An enzyme that hydrolyzes one or more phosphodiester bonds in nucleic acid polymers is called a:

What is the key enzyme used in a polymerase chain reaction (PCR)?

Gene expression microarrays quantify:

The purpose of restriction fragment polymorphism analysis (RFLP)is to:

How does a dideoxynucleotide (ddNTP)that is used in DNA sequencing methods differ from a typical deoxynucleotide (dNTP)present in normal DNA?

False positive results in molecular testing are most likely caused by which one of the following?

In the dideoxy-termination sequencing method,what causes the termination of the newly synthesized DNA strand?

Regarding detection methods,the most common and preferred sequence-specific label for probes used in nucleic acid analysis is a(n)_____ label.

What is the purpose of the labeled oligonucleotide probe used in hybridization assays?

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