DNA (Sanger) sequencing using dideoxy chain termination.
  • A primer is annealed to the target template DNA.
  • A mixture with DNA polymerase, the four dNTPs, and one dideoxynucleotide (such as ddATP), is added.
  • During primer extension, the polymerase occasionally inserts a ddNTP instead of a dNTP, which lacks the 3'-OH needed to attach the next nucleotide, terminating the chain.
  • Gel electrophoresis separates the synthesized fragments that terminate at the dideoxynucleotide.