GSM7734654: mouse RT, R26-SHH, R26P26; Mus musculus; RNA-Seq
Tumor samples cut in small pieces then dissociated 30 min at 37°C in CO2-independent medium (Gibco) + 0,4 g/l of human albumin (Vialebex) with Liberase TL (Roche) 150 mg/ml and DNase 1 (Sigma) 150 mcg/ml. Dissociated cells were then filtered with a 40 mm cell strainer, then washed and resuspended with C02-independent medium + 0,4 g/l of human albumin. Cells were then continuously maintained on ice or at 4°C. In case of lot of blood cells, the Debris removal kit (Miltenyi Biotec) was used according to the manufacturer's protocol. To enrich in tumoral cells (human samples) the Tumor Cell Isolation Kit (Miltenyi Biotec) was used according to the manufactu  rer's protocol. Cells were then resuspended in PBS + BSA 0.04%. Samples were prepared for concentration of 800 cell/mcl. Sample preparations were loaded on a 10x Chromium instrument (10x Genomics) and libraries were prepared using a Single Cell 3' Reagent Kit (V2 chemistry, 10X Genomics) according to the manufacturer's protocol, targeting 1000 