GSM8060251: KPC-tumor-tissue_Normal-VAT-EVs_scRNA-Seq_Rep2; Mus musculus; RNA-Seq
Transfer the chopped pieces to a 50 ml centrifuge tube, add Collagenase Type  II and Collagenase Type IV, add 15 ul CaCl2 (100 mg/ml) solution and 1mg/ml DNase I . Mix well and place on a shaker at 37 ℃ for 20-30 min. After the digestion is complete, take it out and filter it through a 70um sieve and then a 40um sieve into a new 50 ml centrifuge tube, and immediately place the filtered supernatant on ice. Centrifuge at 400g for 4 min. And resuspend the pellet with 5 ml RBC Lysis buffer. 5 ml of 0.04% BSA was used to resuspend, and then determine the cell concentration using a Countess II FL Automated Cell Counter. According to the requirements of 10X GENOMICS on the machine, suck out the corresponding volume of cell suspension. library preparation was performed on  10 × Genomics system.