Extended Data Fig. 1: Nano3P-seq captures non-polyA-tailed and polyA-tailed RNAs.
(a) Tapestation profiles of synthetic RNAs (‘curlcakes’) after being in-vitro transcribed and polyA tailed (pA). Similar profiles were consistently obtained in independent experiments. (b) Tapestation profiles of the input RNA (curlcake mix) for reverse-transcription and cDNA produced after annealing-based or template-switching based (Nano3P-seq) reverse-transcription. (c) IGV snapshots of synthetic RNAs (Curlcake1 and Curlcake2, see Methods) illustrating that Nano3P-seq captures both non-polyadenylated (above) and polyadenylated (below) RNAs. The PolyA tail region is shown in green. (d) Pie chart showing the abundance of different RNA types in Nano3P-seq of mouse nuclear/mitochondria enriched RNA. (e) IGV snapshot of reads mapping to Aldoc gene with polyA tail shown in green. (f) IGV snapshot of reads mapping to Rps3 and Snord15b genes. PolyA tail can be seen in green on the reads mapping to Rps3 mRNA, while it can’t be seen in Snord15b snoRNA. (g) IGV snapshot of reads mapping to Rn7sk miscRNA, which are not expected to contain polyA tails. (h) Scatter plot of the log transformed concentrations (Attomoles/uL) and read counts of sequin transcripts (Pearson R: 0.89, Slope: 0.92). Each dot represents a sequin transcript. (i) Scatter plot of the replicability of the log (read counts) of synthetic sequins using Nano3P-seq, both at per-gene level (left panel, Pearson’s R: 0.99) as well as per-transcript level (right panel, Pearson’s R: 0.98).
