GSM2994875: COLO205.xenograft.combo.24hr.mm10; Homo sapiens; Mus musculus; RNA-Seq
Tissues were minced, enzymatically digested for 10 minutes using 2.5 mg/ml collagenase Type II, 2.5 mg/ml collagenase Type IV (Gibco), and 0.5 mg/ml DNase (Sigma) in PBS / 1% BSA at 37°C, treated with DMEM containing 10% FBS and filtered (70-µm nylon filter; Falcon). Single cell suspensions were subjected to 1X PharMLyse for 4 (tumor) or 10 (heart) minutes, washed and subjected to dead cell removal kit (130-090-101, Miltenyi). The remaining cells were used for single cell sequencing. Tumor single cell suspensions underwent a depletion step using the following FITC conjugated Abs: COLO205 cells were incubated with anti-CD24 (11-0247-42, eBioscience) and anti-E-Cadherin (324104, Biolegend) using 10 µl anti-FITC-beads for 1x10^6 total cells. The remaining cells were used for single cell sequencing. Cellular suspensions (~6000 cells) were loaded on a Chromium Single Cell Instrument (10X Genomics) to generate single cell GEMs. Single-cell RNAseq libraries were prepared using version 1 Chromium Single cell 3' Library, Gel beads & Mutiplex kit (10X Genomics).
GEO Accession
GSM2994875