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Dideoxy Sequencing exam Flashcards

Dideoxy Sequencing exam
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  • Dideoxy sequencing
    A DNA sequencing method that uses dideoxynucleotides (ddNTPs) as chain terminators.
  • Who discovered dideoxy sequencing?
    Frederick Sanger in 1977.
  • ddNTPs
    Dideoxynucleotides used as chain terminators in dideoxy sequencing.
  • What is another name for dideoxy sequencing?
    Sanger sequencing.
  • Template DNA
    The unknown DNA sequence that is to be determined in dideoxy sequencing.
  • What enzyme is essential for dideoxy sequencing?
    DNA polymerase.
  • DNA primers
    Short DNA sequences that anneal to the template strand to initiate DNA synthesis.
  • What are the four normal deoxyribonucleotides used in dideoxy sequencing?
    dATP, dTTP, dGTP, and dCTP.
  • Chain termination PCR
    A PCR reaction that includes ddNTPs to terminate DNA synthesis at specific nucleotides.
  • How many separate reactions are set up in chain termination PCR?
    Four separate reactions, each with a different ddNTP.
  • Gel electrophoresis
    A technique used to separate DNA fragments by size.
  • What is the purpose of gel electrophoresis in dideoxy sequencing?
    To separate DNA fragments by size to determine the DNA sequence.
  • Chromatogram
    A plot generated by a computer to reveal the DNA sequence.
  • How is the DNA sequence read from a gel?
    From bottom to top, across all lanes, to reveal the complementary DNA sequence from 5' to 3'.
  • Complementary base pairing rules
    A pairs with T, and G pairs with C.
  • What is the role of ddNTPs in chain termination PCR?
    To terminate DNA synthesis at specific nucleotides.
  • PCR products
    DNA fragments generated during chain termination PCR.
  • What does the shortest fragment in gel electrophoresis represent?
    The fragment closest to the 5' end of the PCR product.
  • Mystery DNA
    The unknown DNA sequence being determined in dideoxy sequencing.
  • How are the PCR products analyzed to determine the DNA sequence?
    By arranging them based on size and reading the gel from bottom to top.
  • What is the final step in dideoxy sequencing?
    Determining the DNA sequence from the gel or chromatogram.
  • What does a chromatogram reveal?
    The DNA sequence.
  • What is the significance of the 3' hydrogen group in ddNTPs?
    It terminates DNA synthesis.
  • How many DNA primers are needed in dideoxy sequencing?
    Two DNA primers.
  • What is the purpose of separating reactions into different test tubes in chain termination PCR?
    To use different ddNTPs in each reaction.
  • What does the band at the bottom of the gel represent?
    The first nucleotide in the sequence.
  • What is the complementary sequence of a DNA strand with the sequence 5'-ATCG-3'?
    3'-TAGC-5'.
  • What is the role of DNA polymerase in dideoxy sequencing?
    To synthesize DNA by adding nucleotides to the growing strand.
  • What is the main purpose of dideoxy sequencing?
    To determine the sequence of an unknown DNA template.