RNA-Seq of Spumochlamys bryora
We performed whole-culture total RNA extraction by suspending and collecting a culture of the target testate amoebae in sterile 15 ml conical tube. We centrifuged the culture at 4000 x g at 4 C for 5 min to pellet the cells and discarded the supernatant. We extracted the total RNA from the pelleted amoeboid cells using TRIzol reagent (Sigma-Aldrich, St Louis MO) according to the manufacturers protocol (TRI Reagent RNA isolation reagent). synthesized double-stranded cDNA (dscDNA) by following the Smart-seq2 protocol (Piceli et al. 2014). Following the Smart-seq2 protocol, we prepared libraries to be sequenced on Illumina platform using a Nextera XT DNA Library Prep Kit (Illumina, CA).