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sanger sequencing has been the foundation and gold standard of dna sequencing since 1977. while new sequencing technologies have emerged since sanger is still an invaluable tool for all kinds of labs the basic steps of sanger sequencing are simple the reaction involves template dna a primer dna polymerase nucleotides and fluorescently labeled dideoxynucleotides first the dna is denatured and then cooled to allow the primer to bind the dna polymerase enzyme then lengthens the strand ing to the dna template until it randomly places a di-deoxynucleotide the dioxynucleotide is missing the hydroxyl group which allows the next nucleotide to bind so once it is placed the chain is terminated and will not grow any longer the color of the fluorescent dye on the dye deoxynucleotide represents the last base which was added to that fragment this reaction is repeated until the sample contains the fragments of all different lengths each terminated by a fluorescently labeled dideoxynucleotide when the
