The Impact of cDNA Normalization on Long-Read Sequencing of a Complex Transcriptome.

ABSTRACT: Normalization of cDNA is widely used to improve the coverage of rare transcripts in analysis of transcriptomes employing next-generation sequencing. Recently, long-read technology has been emerging as a powerful tool for sequencing and construction of transcriptomes, especially for complex genomes containing highly similar transcripts and transcript-spliced isoforms. Here, we analyzed the transcriptome of sugarcane, a highly polyploidy plant genome, by PacBio isoform sequencing (Iso-Seq) of two different cDNA library preparations, with and without a normalization step. The results demonstrated that, while the two libraries included many of the same transcripts, many longer transcripts were removed, and many new generally shorter transcripts were detected by normalization. For the same inpu

SUBMITTER: Hoang NV 

PROVIDER: S-EPMC6664245 | biostudies-literature | 2019

REPOSITORIES: biostudies-literature