title: GSM8855671: NKCells_TimeCourse_scRNA; Mus musculus; RNA-Seq
library_construction_protocol: Splenic Ly49H+ NK cells (CD3/TCRb/CD19−NK1.1+CD49b+Ly49H+) from uninfected or MCMV-infected WT C57BL/6 mice were sorted as described and stained with barcoded antibodies (Total-Seq B, BioLegend). After hash-staining, NK cells were pooled and the single-cell RNA-seq from these pooled FACS-sorted cell suspensions was performed on a Chromium instrument (10x Genomics) following the user guide manual for 3′ v.3.1. In brief, FACS-sorted cells were washed once with PBS containing 1% bovine serum albumin (BSA) and resuspended in 1× PBS containing 1% BSA to a final concentration of 700–1,300 cells per μl. The viability of cells was above 80%, as confirmed with 0.2% (w/v) Trypan blue staining (Countess II). Cells were captured in droplets. Following reverse transcription and cell barcoding in droplets, emulsions were broken and cDNA-purified using Dynabeads MyOne SILANE (Thermo Fisher, cat. 37002D) followed by PCR amplification as per the manual instructions. Samples were multiplexed together on one lane of 10x Chromium (using Hash Tag Oligonucleotides (HTOs)). Final libraries were sequenced on Illumina NovaSeq6000 S4 platform (R1, 28 cycles; i7, 8 cycles; and R2, 90 cycles).