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Dideoxy Sequencing quiz

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  • What is another name for dideoxy sequencing?
    Dideoxy sequencing is also known as Sanger Sequencing.
  • What molecule is used as an elongation terminator in dideoxy sequencing?
    Dideoxynucleotides (ddNTPs) are used as elongation terminators.
  • Who discovered dideoxy sequencing and in what year?
    Frederick Sanger discovered dideoxy sequencing in 1977.
  • What is the purpose of using an unknown template DNA in dideoxy sequencing?
    The unknown template DNA is used to determine its sequence through the sequencing process.
  • What enzyme is essential for dideoxy sequencing?
    DNA polymerase is essential for dideoxy sequencing.
  • What role do DNA primers play in dideoxy sequencing?
    DNA primers anneal to the template strand to initiate DNA synthesis.
  • Which four normal deoxyribonucleotides are required in dideoxy sequencing?
    DATP, DTTP, DGTP, and DCTP are required.
  • Why are only small amounts of a single ddNTP added to each reaction tube?
    Small amounts of a single ddNTP are added to terminate DNA synthesis at specific nucleotides.
  • What is the key feature of ddNTPs that causes chain termination?
    ddNTPs lack a 3' hydroxyl group, preventing further DNA elongation.
  • How many separate reactions are set up in chain termination PCR for dideoxy sequencing?
    Four separate reactions are set up, each with a different ddNTP.
  • What is produced during chain termination PCR in dideoxy sequencing?
    A variety of DNA fragments of different lengths are produced, each ending with a ddNTP.
  • How are the DNA fragments separated after chain termination PCR?
    The fragments are separated by size using gel electrophoresis.
  • How is the DNA sequence determined from the gel in dideoxy sequencing?
    The gel is read from bottom to top across all lanes to reveal the complementary sequence from 5' to 3'.
  • What rule is used to deduce the sequence of the mystery DNA from the gel results?
    Complementary base pairing rules are used to deduce the sequence.
  • What alternative method can be used to analyze the DNA sequence besides manual gel reading?
    A computer-generated chromatogram can be used to analyze the DNA sequence.