Figure 1: A schematic comparison between the experimental workflows of the DLAF and dUTP methods.
From: Sequencing of first-strand cDNA library reveals full-length transcriptomes
The rRNA-depleted or polyA-enriched RNA is reverse transcribed in the presence of actinomycin D. In DLAF, the double-stranded adaptors with overhangs are ligated to single-stranded cDNA molecules. The forward strands of adaptors containing dU residues are removed by USER, and the libraries are amplified by PCR. In the dUTP method, second-strand cDNA is synthesized in the presence of dUTP and fragmented by sonication, followed by the standard Illumina library preparation procedure and subsequent degradation of dU-containing second strands by USER. Read_1 indicates the reads in the direction of transcription. Read_2 indicates the reads sequenced from the other end of the cDNA molecules.
