Figure 1. Isoform‐specific measurement of transcript decay rates.

1. Outline of MIST‐Seq protocol to measure isoform‐specific decay rates. Exponentially growing cells are shifted to a restrictive temperature that causes transcriptional arrest in the polymerase mutant rpb1‐1. Total RNA is extracted at multiple time points, mixed with equal amounts of S. pombe RNA, and sequenced using 3′ T‐fill to identify transcript 3′ ends at nucleotide resolution.
2. Snapshot of TOM22 isoform stability data. Normalized counts for 3′ isoforms of the gene across three time points (blue) and their corresponding half‐lives (green and purple) are displayed. Only decay rates for 3′ isoforms with reliable quantifications are shown.
3. Log fit (solid lines, deviation in fit in dashed line) to the decreasing number of normalized read counts for the isoforms displayed in (B) in green and purple demonstrates the decay rate of the respective isoforms.