GSM7912864: neoBM; Homo sapiens; RNA-Seq
The tissue was washed with PBS and cut as small as possible. Then, 3 mL of 1640 culture medium was added to a 15 mL centrifuge tube, and 1350 μL of collagenase Ⅰ, 375 μL of collagenase Ⅳ and 180 μL of hyaluronidase were added in sequence and placed into a 37 °C water bath for 30 min with gentle inversion every 5 min. The resulting suspension was filtered through a 70 µm cell strainer to remove larger chunks of undigested tissue. Red blood cell lysis buffer (SAB, R1010) was added to the cells and incubated at 25 °C for 10 min to remove red blood cells. The obtained cell suspension is quality inspected using a cell counter, and samples that pass the quality requirements are labeled for use in experiments. Quality inspection requires cell viability to be greater than 85%, clustering rate to be less than 15%, and nuclear rate to be greater than 70%. Using a single-cell 3' Library and Gel Bead Kit V3.1 (10x Genomics, 1000121) and Chromium Single Cell G Chip Kit (10x Genomics, 1000120) according to Manufacturer's protocol.