Gel Electrophoresis and PCR Quiz

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Questions and Answers

What is the main purpose of Gel Electrophoresis?

To amplify specific DNA segments

To create copies of DNA segments

To separate DNA fragments based on size (correct)

To identify specific DNA sequences

What is the function of DNA primer constructs in PCR?

They identify specific DNA sequences

They separate DNA fragments based on size

They function as primers in PCR reactions (correct)

They amplify DNA segments

What is the role of flanking sequences in PCR?

They amplify DNA segments

They separate DNA fragments based on size

They bracket the DNA sequence of interest (correct)

They function as primers in PCR reactions

What is the function of heat-resistant DNA polymerase in PCR?

It synthesizes new DNA strands during PCR Signup and view all the answers

What is the purpose of denaturing the DNA in a PCR cycle?

To separate the double-stranded target DNA into single strands Signup and view all the answers

What is the main function of agarose gel electrophoresis in DNA analysis?

To separate DNA fragments based on size Signup and view all the answers

What is the purpose of annealing (binding) of primers to ssDNA in a PCR cycle?

To allow the separated strands to anneal to the two primers Signup and view all the answers

What is the main advantage of Polymerase Chain Reaction (PCR)?

It is a very quick, easy, and automated method used to make copies of a specific segment of DNA Signup and view all the answers

What is added to initiate the synthesis of two new chains complementary to the original DNA chains in PCR?

DNA polymerase and deoxyribonucleoside triphosphates Signup and view all the answers

Which end of the primer does DNA polymerase add nucleotides to during PCR?

3′-hydroxyl end Signup and view all the answers

What is the function of Taq antibody in PCR?

Stop the reaction Signup and view all the answers

What is the purpose of using a heat-stable DNA polymerase like Taq polymerase in PCR?

To avoid denaturation and the need for addition at each cycle Signup and view all the answers

Where was the bacterium Thermus aquaticus discovered?

Yellowstone National Park Signup and view all the answers

How many cycles are typically run during the PCR process?

20–30 cycles Signup and view all the answers

What is the outcome of a PCR experiment?

Amplifying the DNA by a million-fold to a billion-fold Signup and view all the answers

What is the function of DNA polymerase in PCR?

Adds nucleotides to the 3′-hydroxyl end of the primer Signup and view all the answers

What is the purpose of denaturing the DNA strands in PCR?

To separate the DNA strands Signup and view all the answers

Study Notes

PCR (Polymerase Chain Reaction)

The main purpose of Gel Electrophoresis is to separate and visualize DNA molecules based on their size and charge.
The function of DNA primer constructs in PCR is to provide a template for DNA synthesis, binding to the target DNA sequence.

PCR Cycle

The function of flanking sequences in PCR is to provide a binding site for primers, allowing for the amplification of a specific DNA sequence.
Heat-resistant DNA polymerase (e.g. Taq polymerase) is used in PCR to synthesize new DNA strands, surviving the high temperatures required for denaturation.
Denaturing the DNA in a PCR cycle is necessary to separate the double-stranded DNA into single strands, allowing for primer binding and subsequent synthesis.

Agarose Gel Electrophoresis

The main function of agarose gel electrophoresis in DNA analysis is to separate, visualize, and purify DNA molecules based on their size.

Primer Function

The purpose of annealing (binding) of primers to ssDNA in a PCR cycle is to provide a template for DNA synthesis, ensuring the specificity of the PCR reaction.

PCR Advantages

The main advantage of Polymerase Chain Reaction (PCR) is its ability to amplify specific DNA sequences, allowing for the detection and analysis of rare or scarce DNA samples.

PCR Mechanism

Primers with nucleotides are added to initiate the synthesis of two new chains complementary to the original DNA chains in PCR.
DNA polymerase adds nucleotides to the 3' end of the primer during PCR.
Taq antibody is not used in PCR; Taq polymerase is a heat-stable DNA polymerase used in PCR.

Thermal Stabilty

A heat-stable DNA polymerase like Taq polymerase is used in PCR to withstand the high temperatures required for denaturation and extension steps.

Thermus aquaticus

The bacterium Thermus aquaticus, from which Taq polymerase is derived, was discovered in hot springs in Yellowstone National Park.

PCR Cycles

Typically, 20-40 cycles are run during the PCR process.

PCR Outcome

The outcome of a PCR experiment is the amplification of specific DNA sequences, allowing for their detection and analysis.

DNA Polymerase

The function of DNA polymerase in PCR is to synthesize new DNA strands by adding nucleotides to the primer, complementary to the target DNA sequence.
Denaturing the DNA strands in PCR is necessary to separate the double-stranded DNA into single strands, allowing for primer binding and subsequent synthesis.

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Description

Test your knowledge of Gel Electrophoresis and PCR with this quiz. Learn about agarose gel electrophoresis, restriction fragment analysis by Southern blotting, and Polymerase Chain Reaction (PCR) technique. Challenge yourself with questions on DNA extraction, isolation, and the importance of these techniques in molecular biology.