Figure 3. Extension of primer PG5-25 using dNTPs or rNTPs as nucleotide substrates by wild-type (WT) RT, and mutants Y115V, A114S, A114G, A114V and A114T.

Reactions were carried out in the presence of 30 nM D2-47/PG5-25. All nucleotides were supplied at a 1 mM concentration each, and the amount of active enzyme in the assay was approx. 15–30 nM. Lanes 1–4 correspond to the analysis of aliquots taken after 15, 30, 60 and 120 min respectively. ‘P’ indicates the position of the 25-mer primer, and ‘F’ stands for the full-length product of 47 nucleotides.