RNA-Seq of tentacle: adult Macrodactyla doreensis
For each organization, total RNA was extracted using the RNeasy Mini kit (Qiao Gen, Duesseldorf, North Rhine Westphalia, Germany). Then, RNA extraction was quantified using a nanodroplet spectrophotometer (Thermo Scientific, Waltham, MA, USA), and RNA integrity number (RIN) values were measured using an Agilent 4200 bioanalyzer (Agilent, Santa Clara, CA, USA). Construct Illumina sequencing library from total RNA using NEBNext Ultra RNA library preparation kit (NEB, Beverly, MA, USA). The sequencing of qualified libraries was performed on the Illumina Novaseq 6000 (Illumina, San Diego, CA, USA) high-throughput sequencing platform. The sequencing strategy used is PE 150. Each sample should contain at least 6 GB of sequencing data. Due to the importance of initial error correction in transcriptome assembly, initial data is screened for quality control. After filtering out these low-quality readings (containing more than 1% non sequencing bases or more than 50% low-quality bases) and adapter sequences, use Trinity with default parameters to reassemble the paired end sequences from scratch into overlapping groups.