Bulk RNA-Seq analysis of  MC38 colon carcinoma cells isolated from Rag2-deficient mice
Bulk RNA-Seq analysis of  MC38 colon carcinoma cells isolated from Rag2-deficient mice
MC38-Bap1-Wildtype and MC38-Bap1-Knockout tumor cells expressing BFP fluorescent protein were injected into Rag2-/- mice subcutaneously. On the 10th day of tumor growth, the tumor cell was dissociated into single cells with Tumor Dissociation Kit (Miltenyi,130-096-730), and red blood cells were removed with ACK lysis buffer (Gibco,492-01). The dead cells in tumor cells were removed with Dead Cell Removal kit (Miltenyi,130-090-101), Then the tumor cells were resuspended with Facs buffer and BFP positive tumor cells were sorted by flow cytometry.  Then RNA was extracted with TRIzol reagent (Ambion 15596-026)follow the instructions. RNA was breaken into smart fragments with fragment buffer, and cDNAs were synthesized using the RNA fragments as templates, after which NEBNext Adaptor with hairpin loop structure were ligated ,and 3 µl USER Enzyme (NEB, USA) was used with purified adaptor-ligated cDNA at 37°C for 15 m in followed by 5 min at 95 °C before PCR. Then PCR was performed with Phusion High -Fidelity DNA polymerase, Universal PCR primers and Index (X) Primer.