GSM8708213: 20181217_2of2; Danio rerio; RNA-Seq
Single-cell whole transcriptome amplification was performed using the Fluidigm C1 Single-Cell Auto Prep System (C1 System) as per the manufacturer's recommendations (full details available at Fluidigm.com). Briefly, tumor was collected and dissociated with collagenase. The cell suspension was filtered through 35um nylon mesh and resuspended at a concentration of 250 cell per μL of complete media, mixed 7:3 with C1 suspension reagent (Fluidigm), and loaded onto C1 Single-Cell Auto Prep chips (C1 chips; Fluidigm). After loading, each of the cell isolation chambers on the C1 chip was optically inspected for the presence of a cell. Single cells was lysed and SMART-Seq (Ramskold et al., 2012) whole transcriptome amplified products were prepared with the C1 System using the SMARTer Ultra Low RNA Kit for Illumina Sequencing (Clontech) and protocols provided by Fluidigm. The whole transcriptome amplification products were harvested from the C1 chip, diluted to a concentration of 0.15 ng/μL, and cDNA libraries were prepared using Nextera XT DNA Sample preparation reagents (Illumina) as per the manufacturer's recommendations.