GSM5580154: GC tissue; Homo sapiens; RNA-Seq
Tumors were obtained after resection in the operating room and were used to prepare single-cell suspensions. Tissue samples were cut into small pieces and transferred to tubes containing 4.7 mL of Dulbecco's modified essential medium, 200 L H, 100 L R, and 25 L AMACS C. MACS™ was separated 3 times, and the sample was incubated at 37 °C for 30 min at each dissociation step interval. After disassembly, the sample was passed through a filter with a diameter of 40 μm, and the suspension was centrifuged at 300 × g for 7 min to completely remove the supernatant. Next, erythrocytes were removed using erythrocyte lysate (10×) (Sigma–Aldrich, St. Louis, Mo, USA). Finally, a dead cell removal kit (Miltenyi Biotec) was used to ensure cell viability >90%. RNA libraries were prepared for sequencing using standard Illumina protocols