Long-read nascent RNA sequencing from S. pombe wild-type and prp2-1 strains

Long-read SMRT cDNA sequencing of nascent RNA from exponentially growing S. pombe 972h- and prp2-1 cells first at 26C and then for 2 hours at 37C was employed to obtain splicing information from single transcripts. Nascent RNA was prepared from the yeast chromatin fraction (Carrillo Oesterreich, Preibisch, Neugebauer, Mol Cell 2010). The nascent 3’ end was labeled with a 3’ DNA adaptor through ligation. The adaptor sequence served as template for full-length reverse transcription and double-stranded cDNA was obtained in a transcriptome-wide PCR. The dataset contains data from 3 replicates per strain. Scripts for demultiplexing the samples can be found on my Github account (https://github.com/lherzel).