RNA-seq of Lingula anatina pedicle 1
RNA was extracted using TRIzol reagent according to the manufacturers instructions. The purified total RNA was then used for library preparation with the Illumina TruSeq Stranded mRNA Library Prep Kit, following the standard protocol. Briefly, mRNA was isolated from 1 g of total RNA using oligo(dT)-coupled magnetic beads and fragmented under elevated temperature. First-strand cDNA synthesis was performed using reverse transcriptase and random primers, followed by second-strand synthesis and 3 adenylation. Adaptors were ligated to the resulting DNA fragments, which were then enriched by PCR and purified using the AMPure XP system (Beckman Coulter). Library quality was assessed using the Qsep400 System (Bioptic) and quantified with a Qubit 2.0 Fluorometer (Thermo Scientific). Sequencing was carried out on an Illumina NovaSeq platform to generate 150 bp paired-end reads.