GSM8962906: Local Patient Derived Organoid 3; Homo sapiens; RNA-Seq
PDOs were mechanical dissociated using a 12 mL syringe and 16-gauge needle followed by the addition of 5 mL of pre-warmed trypsin and incubated for 5 minutes at 37 °C. The addition of trypsin and incubation at 37 °C was repeated 3 additional times. 10% FBS (Gibco) was added and the sample and spun down at 1240 rcf for 5 minutes. Samples were washed twice with 1X PBS (Gibco) to remove residual trypsin. Cells were then stained with trypan blue (Gibco) and counted on a hemocytometer to access cell quantity and viability. Samples were run using the 3' Chromium Single Cell 3' Reagent Kit (10X genomics) and 10, 000 cell recovery was targeted. Manufactures recommendations and instructions were used in constructing the library.