Sanger DNA Sequencing: Basics, Process, and Applications

Sanger sequencing works by incorporating small amounts of chain-terminating nucleotides during DNA replication. These special nucleotides lack a 3' hydroxyl group, which is necessary for forming a phosphodiester bond with the next nucleotide, thus stopping DNA synthesis once incorporated. The method uses four different fluorescently labeled dideoxynucleotides, allowing each termination point to be identified by its unique fluorescent signal.