2k Mouse H2Kb OT-1 Splenocytes (BEAM-T)

Splenocytes from H2Kb-restricted OT-1 transgenic mice (Jackson Laboratory) with specificity for SIINFEKL were labeled with a single peptide (plus an H2Kb control peptide). They were then sorted for CD8+/PE+ T cells as described in the Sample Prep User Guide for Reagent Assembly, Sample Labeling & Flow Sorting for Barcode Enabled Antigen Mapping (CG000595, Rev A).

Peptides:

• OT-1: SIINFEKL (BEAM Conjugate 13)
• H2Kb negative control peptide (BEAM Conjugate 14)

Gene Expression, Antigen Capture (BEAM-T), and VDJ-T amplified libraries were generated from the sorted cells as described in the Chromium Next GEM Single Cell 5' Reagent Kits v2 User Guide (CG000591, Rev A) and sequenced on an Illumina NovaSeq 6000.

Paired-end, dual indexing libraries were sequenced to the following depths:

• GEX: 89,097 mean reads per cell
• Antigen Capture (BEAM-T): 14,725 mean reads per cell
• VDJ-T: 29,665 mean reads per cell

Sequencing read configuration:

• Read 1: 26 cycles (16 bp barcode, 10 bp UMI)
• i5 index: 10 cycles (sample index)
• i7 index: 10 cycles (sample index)
• Read 2: 90 cycles (transcript)

Sequencing data were processed using Cell Ranger v7.1, with web summary and output files available in relevant tabs.

• Cells detected: 1,719
• Median genes per cell: 2,474
• Median UMIs per cell: 9,204

This dataset is licensed under the Creative Commons Attribution license.