Quiz | Real-Time PCR | Attempts: 528 - Trivia , Flashcards & Questions

2.

Nucleotide triphosphates (dNTPs) are added to the growing DNA strand during the _______________ phase.

Extension/elongation
Annealing
Denaturation/separation
Preparation

Correct Answer

A. Extension/elongation

Explanation

During the extension/elongation phase of DNA replication, nucleotide triphosphates (dNTPs) are added to the growing DNA strand. This is when the DNA polymerase enzyme catalyzes the formation of phosphodiester bonds between the incoming nucleotides and the existing DNA strand, resulting in the elongation of the DNA molecule. This process ensures the accurate replication of the genetic information contained within the DNA molecule.

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3.

What does the acronym PCR stand for?

Polymerase Chain Reaction
Polymerase Copying Reaction
Polymerase Copied Repeats
Polymerase Chain Restoration

Correct Answer

A. Polymerase Chain Reaction

Explanation

PCR stands for Polymerase Chain Reaction. This technique is used to amplify a specific segment of DNA through a series of heating and cooling cycles. It involves the use of a DNA polymerase enzyme to copy and replicate the DNA region of interest. The process starts with denaturation, where the DNA strands are separated. Then, primers bind to the target DNA sequence, allowing the DNA polymerase to synthesize new DNA strands. This cycle is repeated multiple times, resulting in the exponential amplification of the desired DNA fragment. PCR is widely used in various applications, including genetic research, diagnostics, and forensic analysis.

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4.

This is the sequence order of RT-PCR.1. Denature2. Anneal3. Detect4. Extend

True
False

Correct Answer

A. False

Explanation

The correct answer is False. The sequence order of RT-PCR is as follows: 1. Denature, 2. Anneal, 3. Extend, and 4. Detect. This is the standard procedure for reverse transcription polymerase chain reaction, where the RNA template is first denatured to separate the strands, then the primers anneal to the target sequence, followed by extension of the primers by a DNA polymerase, and finally the detection of the amplified DNA product.

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5.

Which is/are the property of real-time PCR assays?

Incorporate dyes that bind double-stranded DNA
Incorporate an internal hydrolysis probe
Be performed at single temperature with no specialized instrumentation required
Be interpreted as a plus / minus result or as a quantitative result

Correct Answer(s)

A. Incorporate dyes that bind double-stranded DNA
A. Incorporate an internal hydrolysis probe
A. Be interpreted as a plus / minus result or as a quantitative result

Explanation

Real-time PCR assays incorporate dyes that bind double-stranded DNA, which allows for the detection and quantification of the DNA target. They also incorporate an internal hydrolysis probe, which is a specific sequence of DNA that binds to the target DNA and releases a fluorescent signal during amplification. This probe helps in the accurate detection of the target DNA. Finally, the results of real-time PCR assays can be interpreted as either a plus/minus result (presence or absence of the target DNA) or as a quantitative result (measuring the amount of target DNA present).

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6.

Choose the statement that correctly finishes the sentence:“A PCR reaction that contains only one copy of the target sequence (1 copy /reaction)…

Is typically amplified in a highly repeatable manner”
May amplify but is detection is not likely to be highly repeatable”
Can be precisely and accurately quantified using quantitative PCR”
All of the above

Correct Answer

A. May amplify but is detection is not likely to be highly repeatable”

Explanation

A PCR reaction that contains only one copy of the target sequence may amplify, but its detection is not likely to be highly repeatable. This is because when there is only one copy of the target sequence, there is a higher chance of experimental variability and errors in the detection process. Therefore, although amplification may occur, the results may not be consistently reproducible.

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7.

RNA is copied into complementary DNA (cDNA) by:

Taq DNA polymerase
RNA polymerase II
Reverse transcriptase
Uracil-N-Glycosylase

Correct Answer

A. Reverse transcriptase

Explanation

Reverse transcriptase is an enzyme that is responsible for the synthesis of complementary DNA (cDNA) from an RNA template. It is commonly used in molecular biology techniques such as reverse transcription polymerase chain reaction (RT-PCR) to convert RNA into DNA. This enzyme is derived from retroviruses and is capable of catalyzing the reverse transcription of RNA into DNA. Therefore, it is the correct answer for the given question.

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8.

The reverse transcriptase reaction can be primed by

Target sequence specific primers
Random hexamers
Oligo dT primers
All of the above

Correct Answer

A. All of the above

Explanation

In the reverse transcriptase reaction, primers are used to initiate the synthesis of cDNA from RNA. These primers can be target sequence specific, meaning they are designed to bind to a specific region of the RNA molecule. They can also be random hexamers, which are short DNA sequences that bind randomly to the RNA template. Additionally, oligo dT primers, which are composed of a string of thymine bases, can be used to bind to the poly-A tail found on most mRNA molecules. Therefore, all of the given options - target sequence specific primers, random hexamers, and oligo dT primers - can be used to prime the reverse transcriptase reaction.

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9.

Select the application of RT-PCR.

Drug therapy efficacy
Verification of microarray results
Allelic discrimination assays or SNP genotyping
Quantitative mRNA expression studies
DNA replication efficiency

Correct Answer(s)

A. Drug therapy efficacy
A. Verification of microarray results
A. Allelic discrimination assays or SNP genotyping
A. Quantitative mRNA expression studies

Explanation

RT-PCR (Reverse Transcription Polymerase Chain Reaction) is a technique used to amplify and detect specific RNA molecules. It is commonly used in various applications such as drug therapy efficacy, verification of microarray results, allelic discrimination assays or SNP genotyping, and quantitative mRNA expression studies. In drug therapy efficacy, RT-PCR can be used to determine the effectiveness of a drug by measuring the expression levels of specific genes. In verification of microarray results, RT-PCR can be used to validate the gene expression data obtained from microarray experiments. In allelic discrimination assays or SNP genotyping, RT-PCR can be used to detect specific genetic variations. In quantitative mRNA expression studies, RT-PCR can be used to measure the expression levels of specific mRNA molecules.

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10.

Human errors limits the accuracy of the real-time PCR results:

Incorrect data analysis
improper assay development
Destroy of DNA
Unwarranted conclusions

Correct Answer(s)

A. Incorrect data analysis
A. improper assay development
A. Unwarranted conclusions

Explanation

Human errors can limit the accuracy of real-time PCR results in several ways. Incorrect data analysis can lead to misinterpretation of the results and incorrect conclusions. Improper assay development, such as using incorrect primer sequences or suboptimal reaction conditions, can result in inaccurate measurements. Additionally, unwarranted conclusions can be drawn if proper controls and validation steps are not followed, leading to false interpretations of the data. These human errors highlight the importance of careful and meticulous experimental design, execution, and analysis in real-time PCR to ensure accurate and reliable results.

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Quiz | Real-Time PCR

Who received the Nobel Prize in Chemistry in 1993 for inventing PCR?

Quiz Preview

Nucleotide triphosphates (dNTPs) are added to the growing DNA strand during the _______________ phase.

What does the acronym PCR stand for?

This is the sequence order of RT-PCR.1. Denature2. Anneal3. Detect4. Extend

Which is/are the property of real-time PCR assays?

Choose the statement that correctly finishes the sentence:“A PCR reaction that contains only one copy of the target sequence (1 copy /reaction)…

RNA is copied into complementary DNA (cDNA) by:

The reverse transcriptase reaction can be primed by

Select the application of RT-PCR.

Human errors limits the accuracy of the real-time PCR results: