Question 1

The three basic steps of polymerase chain reaction (PCR)are denaturation, primer annealing, and primer extension. Which step requires DNA polymerase?

Accepted Answer

A) Primer extension 
B) Denaturation 
C) Primer annealing 
D) All of the above 
A) Primer extension 
B) Denaturation 
C) Primer annealing 
D) All of the above

Question 2

Since Taq polymerase cannot function without free magnesium ions (Mg++)and many chemical species can bind Mg++, a gross excess of Mg++ must always be added to any PCR reaction mixture.

Accepted Answer

A) True 
 B)False

Question 3

mRNA preparations that have been treated with diethylpyrocarbonate (DEPC)can usually stored several weeks in the freezer.

Accepted Answer

A) True 
 B)False

Question 4

Explain how reverse transcriptase PCR can be used to accurately quantify RNA in different samples.

Accepted Answer

First, efforts are made to standardize t

Question 5

Why is intact mRNA so hard to purify?

Accepted Answer

A) It is inherently a molecule with a short life. 
B) RNases are relatively resistant to destruction. 
C) RNases are present in almost every environment, including on human skin. 
D) All of the above. 
A) It is inherently a molecule with a short life. 
B) RNases are relatively resistant to destruction. 
C) RNases are present in almost every environment, including on human skin. 
D) All of the above.

Question 6

RNA- dependent DNA polymerase is the proper name for the enzyme ________.

Accepted Answer

reverse tr

Question 7

Describe the principle of TaqMan real- time PCR.

Accepted Answer

In TaqMan real- time PCR, the PCR reacti

Question 8

What technique combines PCR with in situ hybridization?

Accepted Answer

A) Branched chain PCR 
B) In situ amplification 
C) 3' RACE 
D) Nucleic acid sequence based amplification 
E) 5' RACE 
A) Branched chain PCR 
B) In situ amplification 
C) 3' RACE 
D) Nucleic acid sequence based amplification 
E) 5' RACE

Question 9

What advantage does DNA polymerase from Thermus aquaticus (Taq polymerase)have over DNA polymerase from E. coli?

Accepted Answer

A) It synthesizes DNA more accurately with fewer random nucleotide mismatches. 
B) DNA gyrase does not have to be added to the mixture to unwind the DNA prior to synthesis. 
C) It is more heat stable. 
D) It synthesizes DNA more rapidly. 
A) It synthesizes DNA more accurately with fewer random nucleotide mismatches. 
B) DNA gyrase does not have to be added to the mixture to unwind the DNA prior to synthesis. 
C) It is more heat stable. 
D) It synthesizes DNA more rapidly.

Question 10

When making mRNA preparations for reverse transcriptase PCR, it is most important to eliminate contamination from

Accepted Answer

A) transfer RNA. 
B) ribosomal RNA. 
C) nucleolar RNA. 
D) DNA. 
E) All of the above. 
A) transfer RNA. 
B) ribosomal RNA. 
C) nucleolar RNA. 
D) DNA. 
E) All of the above.

Question 11

Even a small amount of contaminating nucleic acid can cause a false positive PCR result.

Accepted Answer

A) True 
 B)False

Question 12

How can one quantify the amount of nucleic acid that is present in a sample and determine its purity?

Accepted Answer

Both DNA and RNA absorb ultraviolet ligh

Question 13

Reverse transcriptase PCR can only be done on pure preparations of messenger RNA as ribosomal RNA and transfer RNA interfere significantly.

Accepted Answer

A) True 
 B)False

Question 14

-A PCR amplification was done on a positive control containing the template DNA, a negative control without the template DNA and two samples. Oligonucleotide primers of 21 base pairs were used, and the expected amplicon product is about 160 base pairs. Observe the agarose gel electrophoresis done on the amplicons, and give the most appropriate interpretations.

Accepted Answer

A) The assay must be repeated because the positive control has more than one amplicon. 
B) The assay must be repeated because the amplicon is not the right size. 
C) The assay must be repeated because there is evidence of "primer dimer" formation. 
D) Sample 1 is positive for the template DNA, and sample 2 is negative for the template DNA. 
E) The assay must be repeated because the negative control is not negative. 
A) The assay must be repeated because the positive control has more than one amplicon. 
B) The assay must be repeated because the amplicon is not the right size. 
C) The assay must be repeated because there is evidence of "primer dimer" formation. 
D) Sample 1 is positive for the template DNA, and sample 2 is negative for the template DNA. 
E) The assay must be repeated because the negative control is not negative.

Question 15

A standard reverse transcriptase PCR assay uses ________(how many?)primer(s)to convert mRNA to cDNA.

Accepted Answer

The answer of A standard reverse transcriptase PCR assay uses...

Question 16

Review the protocol herein and identify its name. Two labeled probes attached to target DNA. DNA polymerase fills in the nucleotides between the p and DNA ligase joins the two pieces. The product is denatured into single strands, and the process i repeated. Amplified target DNA is detected via the labels on the probes.

Accepted Answer

A) Molecular beacon PCR 
B) Ligase chain reaction 
C) 5' RACE PCR 
D) Branched chain PCR 
E) Nucleic Acid Sequence Based Amplification 
A) Molecular beacon PCR 
B) Ligase chain reaction 
C) 5' RACE PCR 
D) Branched chain PCR 
E) Nucleic Acid Sequence Based Amplification

Question 17

Reverse transcriptase was discovered in

Accepted Answer

A) retroviruses. 
B) adenoviruses. 
C) herpes viruses. 
D) Coxsackie viruses. 
E) polioviruses. 
A) retroviruses. 
B) adenoviruses. 
C) herpes viruses. 
D) Coxsackie viruses. 
E) polioviruses.

Question 18

Taq polymerase can add nucleotide bases to a DNA strand at a rate of about

Accepted Answer

A) 10 dNTPs per minute. 
B) 1000 dNTPs per minute. 
C) 100 dNTPs per second. 
D) 10 dNTPs per second. 
E) 100 dNTPs per minute. 
A) 10 dNTPs per minute. 
B) 1000 dNTPs per minute. 
C) 100 dNTPs per second. 
D) 10 dNTPs per second. 
E) 100 dNTPs per minute.

Question 19

Which amplification technique listed is best to use if one wishes to characterize the 5' end of an mRNA transcript?

Accepted Answer

A) 3' RACE 
B) Nucleic acid sequence based amplification (NASB
A)  
C) Branched chain DNA detection 
D) Ligase chain reaction 
E) 5' RACE 
A) 3' RACE 
B) Nucleic acid sequence based amplification (NASB
A)  
C) Branched chain DNA detection 
D) Ligase chain reaction 
E) 5' RACE

Question 20

How does one assure that all PCR amplicons are double- stranded?

Accepted Answer

A) By lengthening the final primer extension step from 1-2 minutes to up to 10 minutes. 
B) By adding DNA ligase to the mixture. 
C) By holding the reaction mixture at annealing temperature for about 5 minutes at the end of the cycle. 
D) By making sure the dNTPs in the mixture are in excess. 
A) By lengthening the final primer extension step from 1-2 minutes to up to 10 minutes. 
B) By adding DNA ligase to the mixture. 
C) By holding the reaction mixture at annealing temperature for about 5 minutes at the end of the cycle. 
D) By making sure the dNTPs in the mixture are in excess.

The three basic steps of polymerase chain reaction (PCR)are denaturation, primer annealing, and primer extension. Which step requires DNA polymerase?

Since Taq polymerase cannot function without free magnesium ions (Mg++)and many chemical species can bind Mg++, a gross excess of Mg++ must always be added to any PCR reaction mixture.

mRNA preparations that have been treated with diethylpyrocarbonate (DEPC)can usually stored several weeks in the freezer.

Explain how reverse transcriptase PCR can be used to accurately quantify RNA in different samples.

Why is intact mRNA so hard to purify?

RNA- dependent DNA polymerase is the proper name for the enzyme ________.

Describe the principle of TaqMan real- time PCR.

What technique combines PCR with in situ hybridization?

What advantage does DNA polymerase from Thermus aquaticus (Taq polymerase)have over DNA polymerase from E. coli?

When making mRNA preparations for reverse transcriptase PCR, it is most important to eliminate contamination from

Even a small amount of contaminating nucleic acid can cause a false positive PCR result.

How can one quantify the amount of nucleic acid that is present in a sample and determine its purity?

Reverse transcriptase PCR can only be done on pure preparations of messenger RNA as ribosomal RNA and transfer RNA interfere significantly.

A standard reverse transcriptase PCR assay uses ________(how many?)primer(s)to convert mRNA to cDNA.

Reverse transcriptase was discovered in

Taq polymerase can add nucleotide bases to a DNA strand at a rate of about

Which amplification technique listed is best to use if one wishes to characterize the 5' end of an mRNA transcript?

How does one assure that all PCR amplicons are double- stranded?

What Is Immunology

Overview of the Immune System

Immunoglobulins

Infection and Immunity: Hostparasite Relationships

The Complement System

Hypersensitivity

Immunologic Detection of Infectious Diseases

Autoimmune Diseases and Immunodeficiency Disorders

Transplant Immunology

Tumor Immunology

Clinical Applications of Flow Cytometry

Hybridization Techniques

Molecular Applications in Cytogenetics

Histocompatibility Laboratory Techniques and Applications

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Exam 13: Amplification Techniques

The three basic steps of polymerase chain reaction (PCR)are denaturation, primer annealing, and primer extension. Which step requires DNA polymerase?

Since Taq polymerase cannot function without free magnesium ions (Mg++)and many chemical species can bind Mg++, a gross excess of Mg++ must always be added to any PCR reaction mixture.

mRNA preparations that have been treated with diethylpyrocarbonate (DEPC)can usually stored several weeks in the freezer.

Explain how reverse transcriptase PCR can be used to accurately quantify RNA in different samples.

Why is intact mRNA so hard to purify?

RNA- dependent DNA polymerase is the proper name for the enzyme ________.

Describe the principle of TaqMan real- time PCR.

What technique combines PCR with in situ hybridization?

What advantage does DNA polymerase from Thermus aquaticus (Taq polymerase)have over DNA polymerase from E. coli?

When making mRNA preparations for reverse transcriptase PCR, it is most important to eliminate contamination from

Even a small amount of contaminating nucleic acid can cause a false positive PCR result.

How can one quantify the amount of nucleic acid that is present in a sample and determine its purity?

Reverse transcriptase PCR can only be done on pure preparations of messenger RNA as ribosomal RNA and transfer RNA interfere significantly.

A standard reverse transcriptase PCR assay uses ________(how many?)primer(s)to convert mRNA to cDNA.

Reverse transcriptase was discovered in

Taq polymerase can add nucleotide bases to a DNA strand at a rate of about

Which amplification technique listed is best to use if one wishes to characterize the 5' end of an mRNA transcript?

How does one assure that all PCR amplicons are double- stranded?

What Is Immunology

Overview of the Immune System

Immunoglobulins

Infection and Immunity: Hostparasite Relationships

The Complement System

Hypersensitivity

Immunologic Detection of Infectious Diseases

Autoimmune Diseases and Immunodeficiency Disorders

Transplant Immunology

Tumor Immunology

Clinical Applications of Flow Cytometry

Hybridization Techniques

Molecular Applications in Cytogenetics

Histocompatibility Laboratory Techniques and Applications

Filters