title: scRNA-seq of mantis shrimp: adult male compound eye
design: Load the cell-nuclear suspension to 10 Chromium to capture cell-nuclear by referring to the manufacturers instructions of 10 Genomics Chromium Single-Cell 3 kit (10 Genomics). Gel beads containing barcode information were bind to the mixture of cell-nuclears and enzymes and then coated with surfactant-water-in-oil droplets located in the microfluidic system to form Gel Beads-In-Emulsions (GEMs). The gel beads in the GEMs that flow into the reservoir and are collected dissolve to release mRNA and reverse transcribes the cDNA with barcode. After the liquid reservoir was destroyed, cDNA amplification reaction was carried out. All GEM products were purified and mixed, and library construction was carried out after passing quality inspection. The cDNA was digestion to form fragments of 200-300 bp, and then performed the end repair, and added the A-tailing and ligation. The target fragments were recovered for quality inspection and quantification to complete the library construction. The constructed library structure from 5 to 3 end was Illumina P5 ligation, P5 sequencing primer, barcode, unique molecular identifier (UMI), poly dT reverse transcription primer, cDNA fragment, P7 sequencing primer, sample index and Illumina P7 ligation. Subsequently, the constructed Chromium 10 Genomics library was sequenced on the Illumina NovaSeq6000 sequencing system (paired-end multiplexing run, 150 bp) at a minimum depth of 20,000 reads per cell.