RNA-Seq of Difflugia cf. capreolata
A single-cell was isolated directly from an environmental sample. The cell was cleaned through sequential dilutions in autoclaved filtered Deer Park spring water and left in petri dishes containing autoclaved filtered Deer Park spring water to starve overnight prior to single-cell RNA extraction. We extracted RNA from single cells and synthesized double-stranded cDNA (dscDNA) using a modified version of Smart-seq2 protocol (Piceli et al. 2014) that includes additional freeze-thaw steps to improve cell lysis as described in Onsbring et al. (2020). Following the Smart-seq2 protocol, we prepared libraries to be sequenced on Illumina platform using a Nextera XT DNA Library Prep Kit (Illumina, CA).