2.4 RNA extraction and sequencing
To estimate the coverage rate of the
assembled genome over gene regions and to predict gene models, RNA-seq
was performed to generate transcript data for multiple tissues,
including the brain, muscle, liver, spleen, olfactory organ, retina, and
gonad tissues collected from the same individual. RNA was extracted
separately using Trizol Reagent (Invitrogen, Camarillo, USA), and RNA
quality was checked using a Nanodrop spectrophotometer (Labtech,
Ringmer, UK). The RNA was used to construct the Illumina RNA-seq library
as described in a previous study (Zhu et al. 2014). These
transcript libraries were sequenced using an Illumina NovaSeq 6000 in
PE150 mode (Illumina Inc., San Diego, CA, USA).