The dideoxynucleotide chain terminator method also known as the Sanger method, uses DNA polymerase and mixes of the four deoxynucloeitdes with one of four of dideoxynucleotides (ddNTPs) for generating a DNA fragments terminated at any specific nucleotides. It requires a single-stranded DNA molecules as a template, a DNA polymerase, DNA primer, normal deoxynucleotides (dNTPs), and ddNTPs that that essentially terminate DNA strand elongation. A primer is annealed to a specific region on the template DNA strand, which provides a starting point to synthesize new DNA strand in the presence of DNA polymerases. The ddNTPs absence the 3’-hydroxyl group of dNTP, which is required for the phosphodiester bond formation between one nucleotide to the following
