Extended Data Fig. 1: Comparison of RPF-based methods with different library construction strategies.

(a) Schematics showing workflows for different Ribo-seq based methods including the conventional Ribo-seq³², Ligation-free Ribo-seq³⁴ and Ribo-lite. (b) Barcharts showing the marker contamination for 500-cell samples by PAGE purification with RNA (left) or ssDNA (right) markers. Results are from two (for RNA marker) and three (for ssDNA marker) independent experiments. (c) A table showing the RPF-based methods and datasets that are used for comparison, including two conventional Ribo-seq^32,37, Ligation-free Ribo-seq³⁴ and Ribo-lite. (d) Bar plots showing the percentages of reads that are adapter, rRNA or clean reads (non-rRNA mapped reads) in different datasets. Results are from two independent experiments. (e) Bar plots showing the metagene read distribution of the 5′-end of reads from Ribo-seq and mRNA-seq data around the translation start codon and stop codon. Read position relative to ORFs (0, 1, 2) are shown in different colors. The schematic shows the ribosome binding around start/stop codon. Results are from two independent experiments.