GSM8698012: PBMC and Thyroid Control  patients3,4; Homo sapiens; RNA-Seq
Peripheral blood mononuclear cells (PBMC) were isolated by density gradient centrifugation. Intrathyroidal blood mononuclear cells were also isolated by density gradient centrifugation , using surgically excised thyroid tissue. They were resuspended in 650 μL of cold sample buffer for loading on a BD Rhapsody cartridge. Targeted scRNA-seq with BCR analysis was performed using the BD Rhapsody Express system (BD Biosciences, NJ, USA). Cell capture and library preparation were performed using the BD Rhapsody Targeted mRNA and AbSeq Reagent Kit (BD Biosciences). Briefly, cells from each sample were captured in a microwell plate with beads. This was followed by cell lysis, bead retrieval, cDNA synthesis, template switching, Klenow extension, and library preparation (targeted gene library using human B-cell expression panel and BCR gene library) following the BD Rhapsody VDJ CDR3 protocol. The final pooled libraries were spiked with 20% PhiX control DNA to increase the sequence complexity and subsequently sequenced (75 bp × 225 bp paired-end) on a HiSeq X Ten/ Illumina NovaSeq X sequencer (Illumina, San Diego, CA).