For this method to work effectively, it requires the following: a DNA primer, a single –stranded DNA template, radioactively or fluorescently labelled nucleotides, a DNA polymerase, and modified nucleotides that play a role in terminating the DNA strand elongation.
The DNA sample is divided into four separate sequencing reactions, containing the DNA polymerase and all four standard deoxynucleotides ( dGTP, dATP, dTTP,dCTP)
Only one of the four dideoxynucleotide (ddGTP, ddATP, ddTTP, ddCTP) is added to each sequencing reaction. The dideoxynucleotides are the chain terminating nucleotides and they lack 3’-OH group necessary for the