2.2 | Nucleic acid isolation, library construction and
sequencing
We firstly removed the intestines of adult individuals under an inverted
microscope to prevent intestinal microorganism contamination. Then,
total genomic DNA was isolated from them using a sodium-dodecyl
sulphate/proteinase K digestion(Gasser et al., 2006) followed by
phenol-chloroform extraction and ethanol precipitation. For PacBio
sequencing, 20-Kb SMRTbell
libraries were constructed using
the SMRTbell Template Prep Kit 1.0 (Pacific Biosciences, USA) and the
SMRTbell Damage Repair Kit (Pacific Biosciences), according to the
manufacturer’ s instructions. The Pacbio sequencing was performed with 1
SMRT cell on the PacBio SMRTplatform. For genome resequencing, 100-bp
pair-end libraries were constructed and sequenced a whole genome
sequence (WGS) short reads on DNBSEQ-T1 platform. For RNA-seq, RNA was
isolated separately from an adult female and a male individual ofB. schroederi using TRlzol reagent (Invitrogen, USA) according to
the manufacturer’s instructions and RNA yields were estimated
spectrophotometrically (NanoDrop 1000), and integrity was verified using
a Bioanalyzer. A total of 22Gb raw fastq data of female and 27 Gb raw
fastq data of male were obtained. All transcripts were used to assess
the completeness of the genome assembly and assist genome annotation.