Illumina NovaSeq 6000 paired end sequencing; Single cell RNA sequencing of three IDHwt glioblastoma patient tumors
Single cell RNA sequencing of three IDHwt glioblastoma patient tumors
Cell dissociation and tissue processing: Fresh tumor samples were dissociated using the gentleMACS Octo Dissociator and Brain tumor dissociation kit (Miltenyi). Myelin and red blood cells were removed using the MACS Myelin removal beads II and ACK Red Blood Cell lysis buffer, respectively. Dead cells were removed (MACS Dead cell removal microbeads) and viable cells were counted by Trypan blue exclusion. Fresh tumor samples were dissociated using the gentleMACS Octo Dissociator and Brain tumor dissociation kit (Miltenyi). Myelin and red blood cells were removed using the MACS Myelin removal beads II and ACK Red Blood Cell lysis buffer, respectively. Dead cells were removed (MACS Dead cell removal microbeads) and viable cells were counted by Trypan blue exclusion. Dissociated tumor cells were processed using the 10x Genomics Chromium Controller and the Chromium Single Cell 3′V2 Library & Gel Bead Kit following the manufacturer's protocols (https://tinyurl.com/ybpg2pfz).  Dissociated tumor cells were processed using the 10x Genomics Chromium Controller and the Chromium Single Cell 3′V2 Library & Gel Bead Kit following the manufacturer's protocols (https://tinyurl.com/ybpg2pfz).