Improving the diversity of captured full-length isoforms using a normalized single-molecule RNA-sequencing method.

ABSTRACT: Human genes form a large variety of isoforms after transcription, encoding distinct transcripts to exert different functions. Single-molecule RNA sequencing facilitates accurate identification of the isoforms by extending nucleotide read length significantly. However, the gene or isoform diversity is lowly represented by the mRNA molecules captured by single-molecule RNA sequencing. Here, we show that a cDNA normalization procedure before the library preparation for PacBio RS II sequencing captures 3.2-6.0 fold more full-length high-quality isoform species for different human samples, as compared to the non-normalized capture procedure. Many lowly expressed, functionally important isoforms can be detected. In addition, normalized PacBio RNA sequencing also resolves more allele-specific hap

SUBMITTER: Hu Y 

PROVIDER: S-EPMC7393167 | biostudies-literature | 2020 Jul

REPOSITORIES: biostudies-literature