GSM8037372: SMARCB1-neg_PTCL-NOS_3; Homo sapiens; RNA-Seq
Isolation of cells for Chromium Fixed RNA profiling was done according to protocol CG000632_RevB (10X Genomics). Briefly, after deparaffiniz­ation and rehydration the tissue sections were dissociated using a heated gentleMACS Octo dissociator for FFPE tissue (130-096-427; Miltenyi Biotec) with freshly prepared Dissociation Enzyme Mix containing Liberase TH (5401151001; Millipore Sigma). After sample filtration, single-nuclei suspensions were counted using Invitrogen Countess 3 Automated Cell Counter (16842556; Thermo Fisher Scientific). To generate fixed RNA gene expression libraries, we used the Chromium Next GEM Single Cell Fixed RNA Sample Preparation Kit, 16 rxn (PN-1000414). Single-nuclei suspensions were mixed with human transcriptome probes and hybridized for 20 h at 42°C and then further processed according to protocol CG000527_RevE (10X Genomics). One million cells per sample were used for probe hybridization, with an expected recovery of 80,000 cells per library (5,000 cells per individual barcode in a 16-plex library).