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. 2022 Apr 29;10:873906. doi: 10.3389/fbioe.2022.873906

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Copyright © 2022 Krebs, Rakotoarinoro, Stech, Zemella and Kubick.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Influence of orthogonal component concentration on fluorescence of RFP-X-GFP reporter protein in the translation mix (TM) and SDS-PAGE of corresponding supernatant fractions SN1 and SN2. (A) Variation of aaRS concentration with 5 µM tRNA and 2 mM ncAA, (B) variation of tRNA concentration with 3 µM aaRS and 2 mM ncAA, (C) variation of ncAA concentration with 3 µM aaRS and 5 µM tRNA for the translation mix (TM) in the FAM channel, the Texas Red channel, ratio of RFU (FAM/TX Red), and the corresponding visualization of the SN1 and SN2 fraction in a radiography of 10% SDS-PAGE (from left to right). Mean values of triplicates were background fluorescence subtracted and are plotted with error bars representing their standard deviation. Green and red arrows depict the full length protein and the termination product, respectively.