Supplementary Figure 2: Illumina sequencing accurately detected U-tags of multiple lengths.
From: Protein-RNA networks revealed through covalent RNA marks
a) Synthetic libraries with various length U-tags, shown here as the reverse complement for clarity. The indicated libraries were paired-end sequenced on an Illumina HiSeq2500. The purple sequence represents the Illumina 5ʹ adapter, the blue sequence represents a poly(A) tail of 12 nucleotides, the red sequence represents U-tags of multiple lengths, and the black sequence represents the U-select RT primer. The starred (*) position in the U0 library was further analyzed in panel b. b) Accuracy of identifying Tagged RNAs by a single non-templated uridine. The percent nucleotide composition of position 13 in Read 2 of the U0 library, which corresponds to the starred (*) position in panel a, was calculated and plotted (n = 310,745). The actual bases detected by sequencing were reverse complemented here for clarity.
