GSM9137075: A673 hybrid rep2; Homo sapiens; RNA-Seq
Hybrid or monoculture spheroids were digested into single cells using trypsin. GFP positive Ewing sarcoma cells were sorted using BD Symphony S6 (CWRU Cytometry & Imaging Microscopy Core), and subject to total RNA extraction using TRIzol (Thermo Fisher, 15596026) according to the manufacturer's instructions. 250ng of total RNA was used to construct RNA-seq library using the NEBNext UltraExpress RNA Library Prep Kit (NEB #E3330) following manufacturer's instructions. Briefly, RNA was polyA enriched using NEBNext Poly(A) mRNA Magnetic Isolation Module (NEB #E7490) and enzymatically fragmented. cDNA was synthesized followed by end repair and adapter ligation, and PCR enrichment with indexed primers. Library was pair end sequenced for 76bp using NovaSeq platform.