TWIST CVRP of Cimex sp.
Sequencing libraries were generated from supernatants of vero cell isolated Stricticimex Coltivirus using the NEBNext Ultra II FS DNA Library Prep Kit for Illumina (New England Biolabs, MA, USA) following the manufacturer's instructions. First-strand cDNA synthesis was performed using the ProtoScript II First Strand cDNA Synthesis Kit (New England Biolabs, MA, USA), followed by second-strand synthesis with the NEBNext Ultra II Non-Directional RNA Second Strand Synthesis Module (New England Biolabs, MA, USA). The resulting double-stranded DNA was fragmented to an average size of approximately 400 bp before adapter ligation. Adapter-ligated DNA fragments were then amplified by PCR using the NEBNext Multiplex Oligos for Illumina. At each step, purification was performed using AMPure XP beads (Beckman Coulter), and DNA quantification was carried out with a Qubit 4.0 fluorometer. Final libraries were sequenced on an Illumina NovaSeq X platform (Macrogen, Inc.), generating paired-end 150 bp (PE150) reads.