GSM7864381: CD45+ DRG cells, GEX; Mus musculus; RNA-Seq
Mice were perfused with ice- cold PBS and neural tissues dissected into cold PBS. On average 40 pooled cervical, thoracic and lumbar DRGs were used. The tissues were digested in 2mg/ml Collagenase I (Gibco 17100-017), 5mg/ml Dispase II (Sigma, D4693) and 0.5mg/ml DNAse I (Roche, 11284932001) at 37C for 30min (Brain) or 40min DRGs. Myelin was removed using 38% Percoll (Sigma, GE17-0891-02). Single cell suspensions of DRG cells from 3 male and 3 female BALB/cAnNRj mice were blocked with FcR blocking solution (Miltenyi Biotec l No.130-092-575) and stained with anti-CD45:PE (BioLegend 103105). Additionally, TotalSeq™ anti-mouse Hashtag antibodies were used to label individual samples The samples were strained  through a 35-micron filter. DAPI was added to the cell solution in order to exclude dead cells. CD45+DAPI− cells were sorted on a BD Influx sorter. The sorted cells were pooled into a single lane on the 10x Genomics Chromium Single Cell 3 ′ v3 system. Library preparation and sequencing were performed at the SciLifeLab sequencing facility, Solna using an Illumina HiSeq  at 61,006 reads/cell. scRNA seq with hashtags