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. 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.