title: GSM8583584: NTC tumor DC1; Mus musculus; RNA-Seq
library_construction_protocol: Tumors were taken at the time of experiment termination and digested in RPMI with collagenase (1 g/100mL HBSS, 10x stock solution, Sigma- Aldrich, C-5138) and DNase (20,000 U/100mL HBSS, 10x stock solution, Sigma-Aldrich, D-5025) at 37°C at 250 rpm in 15 mL conical tubes for 15 minutes. Tumors were processed through a 40 mM cell strainer (Corning, 431750) and CD11c selection was performed using FACS sorting (Sony DH800 Sorter). Selected cells were washed with PBS and lysed with RLT PLUS (QIAGEN, 1053393) with b-mercaptoethanol (VWR, VWRV0482-250ML). RNA was isolated using the RNAeasy Plus Micro kit (QIAGEN, 74034) per the manufacturer's instructions. NEBNext Ultra™ II RNA Library Prep Kit Messenger RNA was purified from total RNA using poly-T oligo-attached magnetic beads. After fragmentation, the first strand cDNA was synthesized using random hexamer primers followed by the second strand cDNA synthesis. The library was ready after end repair, A-tailing, adapter ligation, size selection, amplification, and purification.
identifiers.namespace: GEO
identifiers.id: GSM8583584_r1