RNA-Seq of TCMK-1
Total RNA was extracted using Trizol reagent (Invitrogen) and assessed for quality with an Agilent 2100 Bioanalyzer. For eukaryotic samples, mRNA was enriched with Oligo(dT) beads; for prokaryotic samples, rRNA was removed using the Ribo-Zero Magnetic Kit (Epicentre). The enriched mRNA was fragmented, reverse-transcribed into cDNA with NEBNext Ultra RNA Library Prep Kit (NEB), and processed for end repair, A-tailing, and adapter ligation. The library was purified, size-selected, and PCR-amplified. Sequencing was performed on an Illumina Novaseq6000 by Gene Denovo Biotechnology Co. (Guangzhou, China).