scRNA-seq of homo sapien: GBM stem-like cells PSTAT-treated day4 repB - SN503: Sample S8
Single-nucleus RNA sequencing was performed using the 10X Chromium Controller system and the NextSeq 500. Samples were loaded on a 10X Chromium chip A using single cell RNA-seq 3' v2 chemistry (Single Cell 3' Reagent Kits v2 User Guide Rev D) with a targeted cell recovery of 1,000 cells per time point. Gel emulsions were recovered from the microfluidic chip and full-length cDNA libraries were fragmented, end repaired, A-tailed, and ligated to 10X read 2 adapters prior to SPRI cleanup. Final indexing PCR with P5 and P7 Illumina primers was performed as per standard protocols and libraries were qualified using an Agilent BioAnalyzer 2100 High Sensitivity DNA assay and quantified using the Qubit High Sensitivity DNA kit. They were then pooled in equal amounts, and sequenced on an Illumina NextSeq 500 150bp high output flow cell to a read depth of approximately 50,000 reads per cell.