GSM5217500: COVID_MISA_28; Homo sapiens; RNA-Seq
Blood samples were treated with RBC lysis. 50% of total blood cells were mixed with 50% of enriched lymphycytes (STEM Cell Kit) Libraries were prepared according to 10X Genomics ChromiumTM Single Cell 3' Reagent Guidelines v3.1 Chemistry as per the manufacturer's protocol. In brief, single cells were sorted into 1% BSA–PBS and partitioned into Gel Bead-In-EMulsions (GEMs) using 10xTM GemCodeTM Technology. This process lysed cells and enabled barcoded reverse transcription of RNA, generating full-length cDNA from poly-adenylated mRNA. DynaBeads® MyOneTM Silane magnetic beads were used to remove leftover biochemical reagents, then cDNA was amplified by PCR over 12 cycles. Quality control size gating was used to select cDNA amplicon size prior to library construction. Read 1 primer sequences were added to cDNA during GEM incubation. P5 primers, P7 primers, i7 sample index, and Read 2 primer sequences were added during library construction. Quality control and cDNA quantification was performed using Agilent High Sensitivity DNA Kit. Sequencing was performed using the NovaSeq S2 chip at a read depth of 32,000 reads/cell on an average of 4000 cells/sample.