title: GSM6738033: Hashtagged_scVDJ_sample_2; Homo sapiens; OTHER
library_construction_protocol: Fresh human biospecimens were initially digested into a single cell suspension and cryopreserved in Bambanker Freezing Medium. On the day of the 10X Genomics protocol, cryopreserved samples were thawed, sorted for DAPI- CD45+ CD3+ T cells, hashtagged, stained with TotalSeq C v.1.0 cocktail from Biolegend. Library was prepared using Chromium Single Cell 5ʹ v2 protocol (10x Genomics). Briefly, cDNA amplification included 14-16 cycles and 5.8 ng-20 ng of the material was used to prepare sequencing libraries with 16 cycles of PCR. An aliquot of cDNA generated was used to enrich for V(D)J regions using the Chromium Single Cell V(D)J Enrichment Kit Human T Cell according to the manufacturer's protocol with 10 cycles of PCR during enrichment and 9 cycles during library preparation. Indexed libraries were pooled equimolar and sequenced on a NovaSeq 6000 in a PE150 or PE26/91 run using the NovaSeq 6000 SP or S4 Reagent Kit. An average of 24 million paired reads was generated per sample.