GSM9128546: Pten/Trp53 PX-478 2; Mus musculus; RNA-Seq
Briefly, samples were fixed overnight in Fixation Buffer. After quenching the fixation, samples were dissociated using gentleMACS™ Tissue Dissociator (Miltenyi) and the single-cell suspension was stored at -80°C. After thawing, cell counting was performed using an ADAM MC Plus Fluorescence cell counter (NanoEntek, Seoul, Korea) with a mixed solution of Acridine Orange and 4',6-diamidino-2-phenylindole (DAPI). For each sample, between 506,000 and 780,000 cells were hybridized with a unique Mouse WTA Probe Barcode (10x Genomics, PN-1000492). Single-cell libraries were generated using the Chromium Next GEM Single Cell Fixed RNA reagent kit (10x Genomics, Leiden, The Netherlands), according to the manufacturer's recommendations. cDNA amplification was completed using 8 cycles of PCR. Dual-index library construction was performed with 9 cycles during sample index PCR. Final libraries quantification and quality control were performed using the Bioanalyzer 2100 (Agilent Technologies, Les Ulis, France).