title: GSM7699356: Treg scRNA IL7-Ra antibody tag; Mus musculus; OTHER
library_construction_protocol: Single cell suspensions were prepared from pooled LN from one control (Foxp3-YFP-Cre) and one conditional knockout (Foxp3-YFP-Cre Zfp36l1-fl/fl Zfp36l2-fl/fl) mouse at 14 weeks of age. CD4+ cells were enriched by negative depletion using biotinylated anti-B220, anti-CD8 and anti-F4/80 followed by Streptavidin Dynabeads (Dynal). Treg were sorted to >95% purity as CD4+FR4+YFP+ cells. Cells were labelled using Totalseq oligo-conjugated antibodies (BioLegend) to enable multiplexing of control (hashtag 1: ACCCACCAGTAAGAC) and conditional knockout (hashtag 2: GGTCGAGAGCATTCA) samples, and feature barcoding of CD127 (IL7-Ra; GTGTGAGGCACTCTT). The sorted and labelled single cell suspensions were mixed and loaded onto a Chromium single cell device (10x Genomics) for encapsulation with barcoded gel beads according to the manufacturer's Single Cell 3' v3.1 (Dual Index) protocol. 3'GEX and 3' Feature barcoding libraries were prepared according to the standard manufacturer's protocol. The resulting libraries were sequenced on an Illumina NovaSeq 6000 S1 flowcell