GSM6323617: Immune_K2shIL33KD_2; Mus musculus; RNA-Seq
Single-cell encapsulation and scRNA-seq library prep of FACS-sorted cell suspensions was performed on the Chromium instrument (10x Genomics) following the user manual (Reagent Kit 3' v2). Each sample loaded onto the cartridge contained approximately 5,000 cells at a final dilution of ~500 cells/μl. Transcriptomes of encapsulated cells were barcoded during reverse transcription. cDNA was purified with DynaBeads, followed by amplification per the user manual (Reagent Kit 3' v2). Next, the PCR-amplified product was fragmented, A-tailed, purified with 1.2X SPRI beads, ligated to sequencing adapters and indexed by PCR. Indexed DNA libraries were double-size purified (0.6–0.8X) with SPRI beads.