title: GSM6094425: Glioma, scRNAseq; Homo sapiens; RNA-Seq
library_construction_protocol: The fresh samples of Tumefactive demyelinating lesion and glioma were harvested from the center of lesion and immediately preserved in tissue preservation solution waiting for further processing. Then, the sample was taken out and washed by phosphate-buffered saline (PBS). After that, the sample was minced into small pieces by scissors on ice and enzymatically digested by brain dissociation kit, which contains 1.25 mL of Enzyme N, 2×50 mL of Buffer X (sterile), 1.5 mL of Buffer Y (sterile), 1 vial of Enzyme A (lyophilized powder), and 1 mL of Buffer A. The mixture consisted of sample and enzyme hydrolysate was subjected to the next step for mechanical dissociation via the gentleMACS™ Dissociators. Then, cell clumps and large fragments were strained by a MACS® SmartStrainer (70 μm). After centrifuging cell suspension and resuspending cells with buffer, the final single-cell suspension was generated. According to the manufacturer's protocol, the single-cell suspension was loaded into the Chromium single-cell controller (10X Genomics) for single-cell capturing and downstream library constructions with the Single Cell 3′v2 reagent kit (10x Genomics). Briefly, single cells were suspended and recovered, aiming for 6000 cells. Cells were partitioned into Gel Bead-In-EMulsions (GEMs) with barcoded gel beads. Following cell capture and lysis, complementary DNA was generated and amplified via a S1000TM Touch Thermal Cycler (Bio-Rad).