Tick
RNA extraction, library construction, and sequencing Prior to meta-transcriptomic sequencing, the ticks were categorized and pooled based on collection site, species, gender, host, life stage, and landscape. Total RNA from each pool was extracted and purified using the RNeasy Mini Kit (QIAGEN, Germany) according to the manufacturers instructions. The purified RNA was quantified with a Qubit 4.0 fluorometer, and RNA quality was assessed using an Agilent Bioanalyzer 2200 (Agilent). Next-generation sequencing was applied to each pool, and the sequencing library was constructed utilizing the NEBNext Ultra II Directional RNA Library Prep Kit (NEB, USA). Paired-end transcriptome sequencing (RNA-seq) of the RNA library was conducted on the Illumina NovaSeq 6000 (PE181 bp) sequencing platform (San Diego, USA).