As expected, the percentage of WC interactions with the template calculated for the adjacent residues of the ApAIpA and UpAIpU dinucleotides were much lower, that is, 13.8% and 12.4%. The associated terminal nucleotides of ApAIpA and UpAIpU exhibited negligible WC base pairing. However, as demonstrated for the ApAIpA dinucleotide, the inclusion of complementary (alternative) tU nucleotides within the template increased the population of WC base pairs up to 32.8% for the adjacent and 12.3 % for the terminal nucleotides. The opposite system containing the tUpAIptU dinucleotide is characterised by the lowest population of WC base pairing interactions for both nucleotides, with 8.8 % for the adjacent nucleotide and virtually non formed for the terminal nucleotide. Given that activated tU nucleotides have been generally shown to improve the rate and fidelity of nonenzymatic RNA template copying, the outcome of our final simulation is inconsistent with the experimental observations. Similar to the clustering results, this indicates potential imbalances in the force field parametrization, which is particularly exposed for the terminally bound tUpAIptU dinucleotide.