(A) Long read sequencing of nascent RNA workflow: Nascent RNA molecules are 3’ end ligated and reverse transcribed. A PCR targeting all cDNAs generates double-stranded cDNAs for long read sequencing.
(B) Long read sequencing observations of S. cerevisiae nascent RNA molecules for six representative genes. Presence (thick line) or absence (thin line) of intron sequence and 3’ end position indicate splicing state and Pol II position, respectively.
(C) Long read sequencing observations of S. pombe nascent RNA molecules is represented as described in (B).