TL-seq preferentially recovers capped 5′ ends. (A) Schematic of TL-sequencing (TL-seq). (B) Fragmented RNA (50–80 nt) was treated or mock-treated with pyrophosphatase. Subsequently, both reactions were treated with RNA ligase and a 45-nt adaptor. The gel is overexposed to visualize the shift (bracket). Size markers for a DNA ladder are indicated to the right of the gel. (C) Distribution of where 5′ ends of reads map with and without inclusion of pyrophosphatase. (D) Genes were aligned by their annotated translation start codon and the distribution of reads calculated with or without pyrophosphatase. (E) Comparison of RNA-seq and TL-seq profiles for VMA2. Ordinate is in reads per million reads (rpm); scale bar, lower right.
