Extended Data Fig. 3: OTTR library production and ribosome profiles.
From: Streamlined and sensitive mono- and di-ribosome profiling in yeast and human cells
a, A 0.6X TBE 8% denaturing urea–PAGE gel of OTTR library cDNA generated from human 293 T and yeast cell P1 nuclease RPFs purified by sucrose cushion. The cDNA was directly imaged by the Cy5 dye covalently linked to the ORRT adapter duplex (see Fig. 1a). A parallel OTTR library was synthesized from size marker oligonucleotides (Table 1). Xylene cyanol co-migrates with OTTR adapter dimer cDNA. Horizontal lines indicate excision. b-c, CDS RPF length distribution from yeast (b) and human 293 T (c) RNase I (blue) and P1 nuclease (red) ribosome profiling libraries, as in Extended Data Fig. 1b. d-e, Fraction of sequencing reads mapped to each transcript class yeast (d) and human 293 T (e) ribosome profiling libraries, as in Extended Data Fig. 1a. f, Mean per-base read coverage of cytosolic 18 S and 25 S (or 28 S) rRNA from yeast (left) or human (right) ribosome profiles made from P1 nuclease (red) or RNase I (blue) digestion. Coverage was represented in reads per million total mapped reads (that is, rRNA, tRNA, ncRNA, mRNA, and other genomic loci). g, As in (f) but for 5.8 S and 5 S rRNA coverage. h, As in (f) for mitochondrial rRNA coverage.
