Fig. 5: Single-molecule sensing of total tRNAs from different species.
All measurements were performed as described in Methods. Yeast tRNA was added to cis with a final concentration of 20 ng/μL. E.coli tRNA was added to cis with a final concentration of 2 ng/μL. Trace segmentation and event recognition were performed with the custom machine learning algorithm (Fig. 3a). a A representative trace containing successive yeast tRNA translocation through MspA. Two types of events, termed type 1 (blue triangle) and type 2 (red triangle), were observed, forming the majority of all events that were recorded. Dashed box: Zoomed-in views of representative type 1 and type 2 events, which are respectively marked with i and ii on the trace. The type 1 event has a single blockade level (level 1). The type 2 event contains two blockade levels (level 1 and level 2). b A representative trace containing successive E.coli tRNA translocation through MspA. Two types of events, termed type 1 (blue triangle) and type 2(red triangle) events, were also observed, forming the majority of all events that were recorded. Dashed box: Zoomed-in views of representative type 1 and type 2 events, which are respectively, marked with i and ii on the trace. c The event histogram of blockade amplitude of type 1 and type 2 events (gray: tRNAphe. red: yeast total tRNA. green: E.coli total tRNA). Please note that the current fluctuations between level 1 and 2 show slight variations between events. This variation of fluctuation is more clearly observed in measurements with total tRNAs than those with tRNAphe (Supplementary Fig. 10). However, the \(\% {I}_{b}\) of level 1 and 2 of type 2 events are much more conserved. d Comparison of percentage blockage (\({I}_{p}\)) of type 1 and type 2 events acquired from different tRNA samples. e The proportion of characteristic tRNA events from different sources of tRNAs. Error bars in d and e represent standard deviation, n = 3 independent replicates.
