Single-Cell RNA-Seq of Aposymbiotic Oculina arbuscula
To create single-cell libraries, one symbiotic and one aposymbiotic branch of genotype F of O. arbuscula were sampled for single-cell isolation and 10X cDNA sequencing. Cell isolation samples were analyzed by BUs Single Cell Sequencing Core Facility. The symbiotic sample had a concentration of 2,625 cells/l and viability of 81.6%. The aposymbiotic sample had a concentration of 3,313 cells/l and viability of 86.4%. Libraries were generated following the 10X Genomics Chromium Single Cell 3v3 protocol, and quality was assessed via a Bioanalyzer. Samples were pooled in equimolar concentration and sequenced on an Illumina NextSeq 2000 (P3 100 kit) with a target of 50,000 reads per cell. CellRanger processed reads, which were aligned to concatenated genomes of Astrangia poculata and the algal symbiont Breviolum psygmophilum. Only reads aligning confidently to the host reference were used to generate CellRanger output files.