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Particle picking was performed with gautomatch, all downstream processing was performed in Relion 3.0 beta. Dataset 1 and Dataset 2 were joined after initial 3D classification, where ribosomal particles with P‐site tRNA and SecYEG‐ND were selected from both datasets. Initial processing steps were performed with 2× binned data for stronger signal‐to‐noise ratio at SecYEG‐ND.
