GSM6413350: Sentinel lymph node, OSNA S2; Homo sapiens; RNA-Seq
Total RNA was isolated using RNeasy Plus Mini kit (Quiagen). gDNA decontamination was confirmed. Reverse transcription was performed with 10.5 ng of total RNA using Ion TorrenTM NGS Reverse transcription kit (ThermoFisher Scientific) The Oncomine Immune Response Research Assay – Chef Ready library preparation kit (Thermo Fisher Scientific, Carlsbad, CA, USA) was used for library construction. Automated library preparation, template preparation and subsequent chip loading were performed on the Ion Chef System using the Ion 530 Chef chip kit (ThermoFisher Scientific). For the sequencing step, 25 µl of libraries yielding 50pM were run using the Ion S5 System (ThermoFisher).