Figure 2

Detection of modified RNA bases through the kinetics of SMRT reverse transcription. (a-b) Sequences and example traces of a SMRT reverse transcription trace on m⁶A-containing and unmodified A-containing control template, respectively. Enlarged trace regions show reverse transcription signal for the underlined sequences. (c-d) Distributions of (c) interpulse durations (IPDs) and (d) pulse widths (PWs) for phospholinked T incorporations across m⁶A (orange) and A (blue). The insets in (c) and (d) show histograms of the experimental counts used to determine the cumulative distributions. Phospholinked T binding (k _b ) and dissociation (k _d ) rates were obtained from single exponential fits to the distributions.