title: GSM9240195: scRNA-Seq of Spodoptera exigua hemocytes (naive); Spodoptera exigua; RNA-Seq
library_construction_protocol: three mL of hemolymph was collected from challenged and control larvae on ice and centrifuged at 500 × g for 2 min at 4°C. The hemocyte pellet was washed twice using one mL of cold TC-100 medium (Welgene, Gyeongsan, Korea) supplemented with 10% heat-inactivated (55oC for 30 min) fetal bovine serum (Welgene, Gyeongsan, Korea), which was followed by filtering through a 40 μm cell strainer (Biosharp, China) and centrifugation at 500 × g for 2 min at 4°C. The hemocyte pellet was resuspended using 1x filtered PBS supplemented with 0.04% bovine serum albumin (BSA) (Sigma Aldrich Korea, Seoul, Korea). The number of hemocytes were counted using a hemocytometer. Cell viabilities were assessed by the intensity of the blue fluorescent signal stained by 0.4% trypan blue. Cell suspension samples with viability (>70%) and quantity (>5×10⁵) were subjected to single cell encapsulation. primary data processing were performed by Macrogen Inc. (Seoul, South Korea). Cells were partitioned into Gel Bead-in-Emulsions (GEMs) on the 10x Genomics Chromium Controller using Single Cell 3′ Chemistry v2 kit (10X Genomics, Pleasanton, CA) following the manufacturer's protocol. Within each GEM, a barcoded gel bead bearing a poly(dT) primer captured poly-adenylated mRNA, yielding cell-indexed, UMI-tagged cDNA anchored at the 3′ end. Libraries were prepared according to the Chromium Single Cell 3′ workflow and sequenced on an Illumina NovaSeq 6000 with paired end reads.