CITEseq of human NK: Feature Barcoding library: Sample TML-02-patient-D28_featurebarcode
cDNA was prepared after the GEM generation and barcoding, followed by the GEM-RT reaction and bead cleanup steps.  Purified cDNA was amplified for 11-18 cycles before being cleaned up using SPRIselect beads. The supernatant was then collected for the Cell Surface Protein Library.  Samples were then run on a Bioanalyzer to determine the cDNA concentration for the GEX library preparation.  GEX (Gene Expression) and TotalSeq C Feature libraries were prepared as recommended by the 10x Genomics Chromium Single Cell V(D)J Reagent Kits with Feature Barcode technology for Cell Surface Protein(v1 Chemistry) user guide with appropriate modifications to the PCR cycles based on the calculated cDNA concentration.  For sample preparation on the 10x Genomics platform, the Chromium Single Cell 5' Library and Gel Bead Kit (PN-1000006), Chromium Single Cell A Chip Kit (PN-1000152), Chromium Single Cell 5' Feature Barcode Library Kit, 16 rxns (PN-1000080), Chromium Single Index Kit T for GEX (PN-1000213, Chromium Single Cell Single Index Kit N Set A for Feature (PN – 1000212) were used. The concentration of each library was accurately determined through qPCR utilizing the KAPA library Quantification Kit according to the manufacturer's protocol (KAPA Biosystems/Roche) to produce cluster counts appropriate for the Illumina NovaSeq6000 instrument.  Normalized libraries were sequenced on a NovaSeq6000 S4 Flow Cell using the XP workflow and a 151x8x151 sequencing recipe according to manufacturer protocol.