Figure 4: DLAF detects non-capped 5′ ends of RNA generated by regulatory cleavage.
From: Sequencing of first-strand cDNA library reveals full-length transcriptomes
(a) University of California, Santa Cruz (UCSC) genome browser view of the Mir290 cluster in WT mES cells showing the cleavage events of the primary miRNA (pri-miRNA) during miRNA biogenesis. Green: miRNA-Seq signal. Blue: first sequenced nucleotides of read_1 from the DLAF and dUTP libraries. A distal DLAF peak may represent a previously unknown TSS of pre-miRNA of the Mir290 cluster (green asterisk). Peaks of DLAF read_1-starts precisely match to internal cleavage sites on pri-miRNA (red and brown asterisks). Such peaks were not detected by the dUTP method. (b) Magnified view of two miRNAs, Mir291a and Mir292b (red asterisks). The peaks of DLAF read-starts are located at the nucleotide next to the 3′ end of each miRNA, indicating that DLAF results in the precise detection of 5′ ends of RNA fragments generated during processing of pri-miRNAs. Coverage is normalized to the total non-rRNA and non-mtRNA reads for the dUTP and DLAF libraries.
