Distribution of single-molecule sequencing (SMS) reads among exonic, intronic and intergenic regions in polyA+, RiboMinus and total RNA. RNA samples were prepared as described in the Materals and Methods section of the paper for sequencing. Each source of RNA [K562 cells, human liver or brain tissue and adult flies (Drosophila)] was used either directly (total RNA) or after fractionation by RiboMinus treatment or selection for the polyA containing RNAs. Each sample was sequenced on one or more channels. Reads were aligned to hg18 or dm3 version of the human or fly genomes. After the removal of reads that aligned to the mitochondrial and ribosomal sequences, the remaining sequences were assigned as exonic (white), intronic (grey) or intergenic (black) based on the University of California Santa Cruz genes database and the percentages found for each are shown as pie charts. The exact read data can be found in Additional File 2: Table S1.