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Sanger sequencing reaction mixtures included purified template DNA, a primer, DNA polymerase, and all four dNTPs. Each reaction also included a radiolabeled ddNTP version of just one of the four nucleotides. Only a small amount of each ddNTP was added, however, to ensure that a fraction of the total DNA fragments produced stopped at each occurrence of that base. As with previous methods, separating fragments via length and performing autoradiography allowed scientists to read the final sequence.
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