GSM5835487: DFR1_HTO; Mus musculus; OTHER
GSM5835487
The cell pellet was resuspended and incubated for 30 min on ice with 25μL of staining mix in PBS containing 0.04% BSA, TruStain FcX Block (BioLegend 101320) and the mouse cell surface protein antibody panel containing the following oligo-conjugated anti-mouse antibodies (TotalSeq-A BioLegend) diluted 1:500. Sorted single-cell suspensions were resuspended at an estimated final concentration of 1000 cells/μl and loaded on a Chromium GemCode Single Cell Instrument (10x Genomics) to generate single-cell gel beads-in-emulsion (GEM). Biological replicates (n=4 for each group) were multiplexed using TotalSeq-A Cell Hashing Antibodies. The scRNA/CITE-seq libraries were prepared using the GemCode Single Cell 3' Gel Bead and Library kit, version 3 (10x Genomics 1000128) according to the manufacturer's instructions with the addition of amplification primer (3nM, 5'CCTTGGCACCCGAGAATT*C*C) during cDNA amplification to enrich the TotalSeq-A cell surface protein oligos.