Gel Electrophoresis & PCR Techniques Quiz
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Questions and Answers

What is the purpose of DNA primer constructs in a PCR experiment?

  • To isolate the desired DNA sequence
  • To function as primers in PCR reactions (correct)
  • To cool and anneal the single strands of DNA
  • To separate the double-stranded target DNA
  • In PCR, what is the role of heat-resistant DNA polymerase?

  • To cool and anneal the single strands of DNA
  • To replicate the DNA sequence of interest (correct)
  • To separate the double-stranded target DNA
  • To function as primers in PCR reactions
  • What is the purpose of denaturing the DNA in the PCR cycle?

  • To separate the double-stranded target DNA (correct)
  • To replicate the DNA sequence of interest
  • To cool and anneal the single strands of DNA
  • To function as primers in PCR reactions
  • What is the significance of annealing of primers to single-stranded DNA in PCR?

    <p>To function as primers in PCR reactions</p> Signup and view all the answers

    What is the purpose of having two DNA primer constructs for a PCR experiment?

    <p>To bracket the DNA sequence of interest for amplification</p> Signup and view all the answers

    What is required for a PCR experiment to make copies of a specific segment of DNA?

    <p><strong>Thermocycler</strong></p> Signup and view all the answers

    What is the function of flanking sequences in a PCR experiment?

    <p><strong>To bracket the DNA sequence of interest</strong></p> Signup and view all the answers

    What is the purpose of adding DNA polymerase and deoxyribonucleoside triphosphates in excess to the PCR mixture?

    <p>To initiate the synthesis of two new chains complementary to the original DNA chains</p> Signup and view all the answers

    Which end of the primer does DNA polymerase add nucleotides to during the chain extension process in PCR?

    <p>3′-hydroxyl end</p> Signup and view all the answers

    Why is a heat-stable DNA polymerase (e.g. Taq polymerase) used in PCR?

    <p>To prevent denaturation of the polymerase at high temperatures</p> Signup and view all the answers

    What is the outcome of running 20–30 cycles during the PCR process?

    <p>Amplifying the DNA by a million-fold to a billion-fold</p> Signup and view all the answers

    What is the role of Taq antibody in the PCR process?

    <p>To stop the reaction</p> Signup and view all the answers

    Where was Thermus aquaticus, the bacterium species used for heat-stable DNA polymerase, discovered?

    <p>Yellowstone National Park, USA</p> Signup and view all the answers

    Who discovered the bacterium species Thermus aquaticus?

    <p>Thomas Brock</p> Signup and view all the answers

    What is added at each successive cycle in PCR due to using a heat-stable DNA polymerase?

    <p>Nothing is added at each successive cycle</p> Signup and view all the answers

    Study Notes

    PCR Experiment

    • DNA primer constructs are used to initiate PCR amplification of a specific segment of DNA.
    • Heat-resistant DNA polymerase (e.g. Taq polymerase) is used to synthesize new DNA strands in high-temperature PCR cycles.

    DNA Denaturation

    • Denaturing the DNA in the PCR cycle melts double-stranded DNA into single-stranded DNA, allowing primers to anneal.

    Primer Annealing

    • The purpose of annealing primers to single-stranded DNA is to provide a template for DNA synthesis.
    • Two DNA primer constructs are used to bind to opposite strands of the target DNA segment, enabling amplification.

    PCR Requirements

    • To make copies of a specific segment of DNA, a PCR experiment requires DNA polymerase, deoxyribonucleoside triphosphates, and primers that flank the target sequence.

    Flanking Sequences

    • Flanking sequences are the regions of DNA adjacent to the target sequence, which are amplified along with the target sequence.

    PCR Reaction Mixture

    • Adding DNA polymerase and deoxyribonucleoside triphosphates in excess to the PCR mixture ensures efficient DNA synthesis.

    Chain Extension

    • During the chain extension process, DNA polymerase adds nucleotides to the 3' end of the primer.

    Heat-Stable DNA Polymerase

    • A heat-stable DNA polymerase (e.g. Taq polymerase) is used in PCR to withstand the high temperatures required for DNA denaturation.

    PCR Cycles

    • Running 20-30 cycles during the PCR process results in exponential amplification of the target DNA sequence.

    Taq Antibody

    • There is no role for Taq antibody in the PCR process (Taq refers to the bacterium Thermus aquaticus, which is the source of the heat-stable DNA polymerase).

    Thermus Aquaticus

    • Thermus aquaticus, the bacterium species used for heat-stable DNA polymerase, was discovered in hot springs in Yellowstone National Park.
    • The bacterium was discovered by Thomas Brock and Hudson Freeze in 1969.

    PCR Amplification

    • At each successive cycle, the number of amplified DNA molecules doubles due to the use of a heat-stable DNA polymerase.

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    Description

    Test your knowledge on Gel Electrophoresis, Southern Blotting, and Polymerase Chain Reaction (PCR) techniques used in DNA extraction and analysis. Learn about the importance and applications of these methods.

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