title: GSM8543373: bone marrow, ex vivo, memory T cells, d2, GEX, TCR, CITE-seq.; Homo sapiens; OTHER
library_construction_protocol: Memory T cells were isolated from fresh ex-vivo paired blood and bone marrow samples of three different donors with straight from blood magnetic enrichment (Miltenyi Biotech) Followed by cell sorting trough flow cytometric analysis. Chromium Next GEM Single Cell 5' reagent kits v2 (dual index) with featured barcode technology for cell surface protein (CITE) mapping (10X Genomics) were used as stated in the manufacturer's protocol. Final CITE-Seq libraries were generated after index PCR with dual Index Kit TN Set A (10X Genomics) while final GEX and TCR libraries were generated after fragmentation, adapter ligation and final index PCR with a dual Index Kit TT Set A (10X Genomics). Libraries were quantified using a Qubit HS DNA assay kit (LifeTechnologies) and fragment sizes were determined using a HS NGS Fragment (1-6000 bp) kit (Agilent).All libraries (GEX and CITE and TCR libraries) were sequenced on a NextSeq2000 sequencer (Illumina) using a P3 reagent kit (100 cycles) (Illumina) and following the sequencing conditions recommended by 10x Genomics: read1: 26 nt, read2: 90 nt, index1: 10 nt, index2: 10 nt.(35)