title: GSM9523361: TCR of CD4 T Cells with TET2 KO cultured in vitro at day3 or day6 after 30 cycles of stimulations; Homo sapiens; RNA-Seq
library_construction_protocol: WT and TET2-/- CD4+ T cells were prepared as a single-cell suspension. Cells were tagged with TotalSeq- anti-Human hashtag oligos (DNA barcodes), washed, pooled, and loaded on the 10x Genomics Chromium platform according to the manufacturer's protocol. > Day 200 TET2-/- cells were processed in a separate run using the same workflow. WT and TET2-/- cells collected at days 40 and 90 were labeled with different DNA barcodes using TotalSeq anti-Human Hashtag 1, 2,3, and 4. Single-cell suspensions were processed using the 10x Genomics Chromium Single Cell platform and 10x Genomics TCR V(D)J libraries were constructed using the Chromium Next GEM Single Cell V(D)J v1.1 chemistry kit following the manufacturer's instructions. Libraries sequenced on an Illumina Novaseq6000 (PE101). Cells cultured longer (>200 days were) at day 3 or day 6 after added anti-CD3/anti-CD28 beads were processed in a separate run with the same workflow.