Figure 4.

Graphical plots of yeast tRNA^{Asn (GTT)}, which was used for library preparation, visualized by using the additional module Visualize_V3. The reverse transcription step was performed by using SuperScript^® III Reverse Transcriptase in different reaction buffers. The supplier’s standard reaction buffer (First Strand Synthesis buffer) with Mg²⁺ serves as reference (A), and the tested buffer mixtures differ by increased concentrations of Mn²⁺ [0.5 mM (B), 1.0 mM (C), 3.0 mM (D)] as Mg²⁺ substitute. Sites with error rates of more than 10% are highlighted with yellow arrows, with colored bars indicating the nature of the reads. Mismatch rates are depicted as black crosses, and arrest rates as red lines. The m1A site is located in the middle of the shown sequence segment at position 59.