RNA-Seq of Richtersius coronifer
For transcriptome sequencing and assembly, total RNA was extracted from a single specimen of tardigrade using Direct-zol RNA kit (Zymo Research) and was amplified using SMARTer Ultra Low Input RNA Kit for Sequencing v.3 (Clonetech). Illumina libraries were prepared using KAPA HyperPlus Kit (KAPA BIosystems), and the library was sequenced using a NextSeq 500 High Output Mode 75 cycles kit (Illumina) as single-end 75bp layout. Protocols are detailed in Arakawa et al. (2016) and Yoshida et al. (2018).