chapter 9
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Questions and Answers

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

  • To make billions of copies of a specific DNA sequence (correct)
  • To produce new DNA sequences
  • To identify different types of DNA
  • To analyze the structure of DNA molecules

    • What does 'in vitro' refer to in the context of PCR?

  • DNA modification in a controlled environment
  • DNA replication in a test tube (correct)
  • DNA replication in living cells
  • DNA sequencing in a laboratory setting

    • How does PCR relate to DNA Replication in cells?

  • It is a laboratory version of DNA Replication (correct)
  • It is an alternative process to DNA Replication
  • It is used to inhibit DNA Replication
  • It has no relation to DNA Replication

    • What is one of the applications of PCR mentioned in the text?

    <p>Detecting mutations in specific genes for genetic testing</p> Signup and view all the answers

    How does PCR contribute to forensic analysis?

    <p>Identifying and matching DNA samples from crime scenes to suspects</p> Signup and view all the answers

    In what way does PCR support drug development?

    <p>Identifying and validating drug targets and screening potential drug candidates</p> Signup and view all the answers

    What is the purpose of DNA Extraction Lysis in sample preparation?

    <p>To break cells and release DNA</p> Signup and view all the answers

    What is the role of Proteinase K in the DNA extraction process?

    <p>To dissolve cellular proteins</p> Signup and view all the answers

    What is the final step in DNA purification during sample preparation?

    <p>Eluting the DNA with a slightly alkaline buffer</p> Signup and view all the answers

    What does PCR copy from the sample?

    <p>A very specific sequence of genetic code from a template DNA</p> Signup and view all the answers

    What information is required for the amplification of a specific target sequence using PCR?

    <p>DNA sequence information which flanks the fragment of DNA to be amplified</p> Signup and view all the answers

    What is the role of PCR primers in the amplification process?

    <p>To target a specific sequence of genetic code for amplification</p> Signup and view all the answers

    What does PCR copy from the sample?

    <p>A very specific sequence of genetic code from a template DNA</p> Signup and view all the answers

    What information is required for the amplification of a specific target sequence using PCR?

    <p>DNA sequence information which flanks the fragment of DNA to be amplified</p> Signup and view all the answers

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

    <p>To amplify a specific target sequence of DNA</p> Signup and view all the answers

    What is the purpose of oligonucleotide primers in PCR?

    <p>To initiate DNA synthesis at a specific target sequence</p> Signup and view all the answers

    What is the role of DNA polymerase in PCR?

    <p>Adding nucleotides onto pre-existing 3'-OH groups to extend DNA strands</p> Signup and view all the answers

    Why are two oligonucleotide primers required in PCR amplification?

    <p>To allow amplification of both DNA strands</p> Signup and view all the answers

    Why are two primers required in a PCR reaction?

    <p>To amplify both strands of the double-stranded DNA</p> Signup and view all the answers

    What is the role of the forward primer in PCR?

    <p>It binds to the complementary reverse DNA strand</p> Signup and view all the answers

    What happens if only one primer is used in a PCR reaction?

    <p>Only one strand of the double-stranded DNA will be amplified</p> Signup and view all the answers

    What is the recommended GC content range for a good primer?

    <p>40-60%</p> Signup and view all the answers

    Why is it important for the 3’ of a primer to end in G or C?

    <p>To promote binding</p> Signup and view all the answers

    What impact do G and C bases have on the melting temperature (Tm) of primers?

    <p>They increase Tm</p> Signup and view all the answers

    At what temperature does denaturation of dsDNA usually occur?

    <p>94°C</p> Signup and view all the answers

    How long can denaturation of dsDNA take?

    <p>Up to 2 minutes</p> Signup and view all the answers

    What impact does a higher GC content have on denaturation temperatures?

    <p>Requires higher and longer denaturation temperatures</p> Signup and view all the answers

    What is the recommended annealing temperature (Ta) for PCR primers?

    <p>50-65 °C</p> Signup and view all the answers

    What happens if the annealing temperature in PCR is too high?

    <p>Primers do not anneal efficiently</p> Signup and view all the answers

    How long should the annealing time be maintained ideally in PCR?

    <p>15-60 seconds</p> Signup and view all the answers

    At what temperature can the extension step of PCR usually be carried out?

    <p>70-80°C for 1 to 2 minutes</p> Signup and view all the answers

    What is the optimal operating temperature for Taq polymerase during the extension step for the first 2 kb?

    <p>70-80°C for 1 minute</p> Signup and view all the answers

    How does the required time per additional kb differ between Taq and Pfu polymerases during the extension step?

    <p>Taq requires an additional 1 minute per kb, Pfu requires an additional 2 minutes per kb</p> Signup and view all the answers

    What is the main principle behind Gel Electrophoresis?

    <p>Separation and analysis of macromolecules based on size and charge</p> Signup and view all the answers

    What happens to charged biomolecules when placed in an electric field during Gel Electrophoresis?

    <p>They move toward the opposite charged electrode</p> Signup and view all the answers

    Which direction does DNA move in Gel Electrophoresis?

    <p>Toward the positive electrode</p> Signup and view all the answers

    What happens to negatively charged biomolecules in Gel Electrophoresis?

    <p>They move towards the anode (+)</p> Signup and view all the answers

    What happens to positively charged biomolecules in Gel Electrophoresis?

    <p>They move towards the cathode (-)</p> Signup and view all the answers

    How does size affect the movement of molecules in Gel Electrophoresis?

    <p>Larger molecules will be held up, smaller molecules will be impeded less and move faster</p> Signup and view all the answers

    What is the main role of the gel matrix in gel electrophoresis?

    <p>To serve as a porous medium through which molecules can move during electrophoresis</p> Signup and view all the answers

    What is the purpose of sample loading in gel electrophoresis?

    <p>To load samples containing the molecules to be separated into wells or slots at one end of the gel</p> Signup and view all the answers

    What is the function of the electrophoresis chamber in gel electrophoresis?

    <p>To provide an electric field for molecules within the gel to move toward the oppositely charged electrode</p> Signup and view all the answers

    What type of marker is commonly used to stain the gel after electrophoresis for DNA separation?

    <p>Ethidium bromide</p> Signup and view all the answers

    What information can be obtained by analyzing the separated molecules in the gel after electrophoresis?

    <p>Size, quantity, and purity of the molecules</p> Signup and view all the answers

    For what purpose is gel electrophoresis often used in relation to DNA fragments?

    <p>To determine the fragment size or check the success of DNA amplification</p> Signup and view all the answers

    Who should not take the new drug based on the SNP variation analysis?

    <p>Individuals not carrying the T allele on a specific locus in the CYP2D6 gene</p> Signup and view all the answers

    Which gene's variation analysis is relevant for determining who should not take the new drug?

    <p>CYP2D6 gene</p> Signup and view all the answers

    What did the SNP variation analysis reveal about the new drug's benefit for certain individuals?

    <p>It is not beneficial for individuals not carrying the T allele on a specific locus in the CYP2D6 gene</p> Signup and view all the answers

    What does the electrophoresis gel show?

    <p>The DNA of two male chimpanzees, a female, and an infant</p> Signup and view all the answers

    Why was the DNA from two male chimpanzees tested?

    <p>To determine the paternity of the infant chimpanzee</p> Signup and view all the answers

    What event triggered the DNA testing of the chimpanzees?

    <p>The recent birth of an infant by a female chimpanzee in captivity</p> Signup and view all the answers

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