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Updated: May 5 2017

DNA Sequencing

  • Overview
    • Function
      • determining sequence for a desired DNA sample
    • Process (Sanger/direct method)
      • DNA sample put in 4 different reaction mixtures
        • each contains all 4 deoxynucleotides, 1 different dideoxyneucleotide (ddNTP), and a DNA polymerase
      • dideoxynucleotides stop DNA polymerization at each base
        • generates DNA chains of various lengths that encompass the entire original sequence
      • solutions are separated by gel electrophoresis
      • sequence can be deduced by looking at each electrophoresis row and reading which ddNTP was added to that tube column
    • Clinical use
      • most commonly used when no specific set of mutations is a common cause of disease
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