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Introduction to Polymerase Chain Reaction exam Flashcards

Introduction to Polymerase Chain Reaction exam
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  • PCR
    Polymerase Chain Reaction, a technique used to amplify specific DNA sequences in a test tube.
  • Why is PCR preferred over DNA cloning for quick DNA amplification?
    PCR is quicker and more efficient, taking about 1.5 to 2 hours compared to over 24 hours for DNA cloning.
  • Template DNA
    The DNA sequence of interest that is to be amplified in PCR.
  • What are the four key components required for PCR?
    Template DNA, two complementary primers, thermostable DNA polymerase, and all four deoxyribonucleotides.
  • Primers
    Short DNA sequences that are complementary to the target DNA and serve as starting points for DNA synthesis.
  • Thermostable DNA Polymerase
    An enzyme, often Taq polymerase, that synthesizes new DNA strands and is stable at high temperatures.
  • What is the role of Taq polymerase in PCR?
    Taq polymerase synthesizes new DNA strands during the PCR process.
  • Deoxyribonucleotides
    The building blocks of DNA, including adenine (A), thymine (T), cytosine (C), and guanine (G).
  • How does the number of DNA copies change with each PCR cycle?
    The number of DNA copies doubles each cycle, following the formula 2^n, where n is the cycle number.
  • Amplification
    The process of making many copies of a specific DNA sequence.
  • What is the main advantage of PCR over DNA cloning?
    PCR is much faster and more efficient for generating many identical copies of DNA.
  • Cycle
    A single round of DNA denaturation, primer annealing, and DNA synthesis in PCR.
  • What is the purpose of primers in PCR?
    Primers bind to the target DNA sequences and provide a starting point for DNA synthesis.
  • Denaturation
    The step in PCR where the double-stranded DNA is heated to separate it into two single strands.
  • Annealing
    The step in PCR where primers bind to their complementary sequences on the single-stranded DNA.
  • Extension
    The step in PCR where the DNA polymerase synthesizes new DNA strands by adding nucleotides to the primers.
  • What is the formula to calculate the number of DNA copies after n cycles of PCR?
    2^n, where n is the number of cycles.
  • Why is thermostable DNA polymerase used in PCR?
    It remains stable and active at the high temperatures required for DNA denaturation.
  • What is the role of deoxyribonucleotides in PCR?
    They are the building blocks that the DNA polymerase uses to synthesize new DNA strands.
  • How long does a typical PCR process take?
    Approximately 1.5 to 2 hours.
  • What is the main difference between PCR and DNA cloning?
    PCR occurs in a test tube without living cells, while DNA cloning occurs inside living cells.
  • What is the purpose of the denaturation step in PCR?
    To separate the double-stranded DNA into single strands.
  • What happens during the annealing step of PCR?
    Primers bind to their complementary sequences on the single-stranded DNA.
  • What is the purpose of the extension step in PCR?
    To synthesize new DNA strands by adding nucleotides to the primers.
  • How does PCR help in forensic science?
    It amplifies small amounts of DNA from a crime scene to generate enough material for analysis.
  • What is the significance of the formula 2^n in PCR?
    It represents the exponential increase in the number of DNA copies with each cycle.
  • Why is PCR considered more efficient than DNA cloning?
    PCR can produce many copies of DNA in a short time without the need for cell growth and DNA isolation.
  • What is the role of the template DNA in PCR?
    It provides the sequence that is to be amplified.
  • What are the four deoxyribonucleotides used in PCR?
    Adenine (A), thymine (T), cytosine (C), and guanine (G).