Fig. 4. Translation-uncoupled replication experiments.

a The experiments were performed in two steps. (1) First, one of the host RNA clones (RNA 1, 30 nM) was incubated at 37 °C for 2 h to translate the replicase without RNA replication. (2) The reaction mixture was diluted, and RNA replication was initiated with the translated replicase at 37 °C for 2 h while stopping translation, in the presence or absence of the same concentration (10 nM) of another RNA clone (RNA 2). The replication of each RNA was measured by sequence-specific RT-qPCR. b–e Replication of one or pairs of RNA clones at rounds 0 (b), 120 (c), 155–158 (d), and 228 (e). Colors match those in Fig. 3d. Error bars indicate mean ± SEM (n = 3 or 4 as shown as individual data points). Measurements were taken from distinct samples. Average fold replications (>1.5-fold) were used to draw the directed graphs (Fig. 3d). Source data are provided as a Source data file.