GSM9686225: Fugu spinal cord, C5, scRNA-seq 2; Takifugu obscurus; RNA-Seq
Spinal cord tissue from multiple individuals was pooled per library. Cell dissociation was performed by incubation in pronase solution for 40 min at room temperature, followed by trituration in 100 µL ACSF containing 1% FBS. Volume was adjusted to 3 mL and live cells were enriched by FACS (BD FACSAria III) using propidium iodide (1 µg/mL) exclusion. Cell viability exceeded 80%. Sorted cells were resuspended in 1× DPBS containing 0.04% BSA. Libraries were prepared using the Chromium Single Cell 3' GEM, Library & Gel Bead Kit v3 (10x Genomics). Target cell recovery was 1,000-9,000 cells per replicate depending on tissue availability. Reverse transcription was performed using a C1000 Touch Thermal Cycler (Bio-Rad). Libraries were sequenced on an Illumina HiSeq X platform (paired-end 100 bp reads) targeting an average of 20,000 read pairs per cell.