High-throughput single-molecule long-read RNA sequencing analysis of tissue-specific genes and isoforms in lettuce (Lactuca sativa L.)

This study utilized the HIT-ISOseq method for high-throughput sequencing of RNA isoforms across multiple lettuce samples, generating millions of long reads per PacBio Sequel II SMRT Cell. Analysis of six tissue types revealed tissue-specific gene expression and RNA isoforms, facilitating updates to the lettuce reference genome annotation with expanded functional annotations. Overall design: The experiment involved two samples each of lettuce leaves, roots, and stems. After RNA extraction and cDNA amplification, PacBio sequencing libraries were constructed using the HIT-ISOseq method. Each sample was tagged with a barcode SMRTbell adapter and sequenced in HiFi mode.