Updated: 5/5/2017

DNA Sequencing

Review Topic
  • 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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