GSM8809362: SAOS2, LRRC15+; Homo sapiens; RNA-Seq
For each cell line, cells were FACS sorted for the top 1% LRRC15+ cells by AlexaFluor647-DUNP19 staining with a target cell population of 10,000 viable cells. Cells were resuspended in 1x PBS+0.04% BSA solution and counted for viability before performing the 10X Genomics single cell protocols for library preparation. Single cells were captured within gel bead emulsions (GEM) according to 10X Genomics' Chromium Single Cell platform protocols. Within individual GEM droplets, cells are lysed and barcoded before reverse transcription and cDNA pooling. Post-GEM cleanup was performed with Dynabeads and cDNA amplified, cleaned, and quantified. A 3'GEX library was constructed and quality control performed with Agilent ScreenTape Analysis. Libraries were sequenced with Novaseq S4 (2000-2500 M reads per lane), with a read depth of 20,000.