GSM8324306: HTO library of 5 GFP negative mice; Mus musculus; RNA-Seq
SVF cells were isolated from 5 25-30 months old female P mice. Cells from each donor were labelled with cell hashing antibodies (TotalSeq-A anti-mouse Hashtag, BioLegend) according to the manufacturer's protocol. Cells were then washed, and resuspended in 0.04% BSA/PBS. Labelled cells from each donor were pooled at equal proportions, then immediately sorted on a BD FACSAria Fusion cell sorter (BD Biosciences) for viable GFPhigh and non-GFPhigh cell populations. Cells from wildtype mice were used as GFP background for gating and sorting. Cell viability was assessed on a Countess II automated cell counter (Thermo Fisher Scientific). Up to 12,000 cells were loaded onto one lane of a 10X Chromium X. Single cell capture, barcoding and library preparation were performed using the 10X Chromium platform version 3.1 chemistry according to the manufacturer's protocol (CG000388). cDNA and libraries were checked for quality on Agilent 4200 TapeStation (Agilent Technologies, Santa Clara, CA) and Qubit Fluorometer (Thermo Fisher Scientific), quantified by KAPA qPCR. Library was sequenced on an Illumina NovaSeq 6000 S4 v1.5 200 cycle flow cell lane, targeting 6,000 barcoded cells with an average sequencing depth of 50,000 reads per cell. 10X Chromium platform version 3.1 chemistry according to the manufacturer's protocol (CG000388)