GSM8809605: nucleus pulposus cells treated with NPPC_Exo_2; Rattus norvegicus; RNA-Seq
Total RNA from NPCs was isolated using a TRIzol reagent (Invitrogen, U.S.A.). The cleaved RNA fragments were reversetranscribed to create the cDNA by SuperScript™ II Reverse Transcriptase (Invitrogen, cat. 1896649, USA), which were next used to synthesise U-labeled second-stranded DNAs with E. coli DNA polymerase I (NEB, cat.m0209, USA), RNase H (NEB, cat.m0297, USA) and dUTP Solution (Thermo Fisher, cat.R0133, USA）. An A-base is then added to the blunt ends of each strand, preparing them for ligation to the indexed adapters. Each adapter contains a Tbase overhang for ligating the adapter to the A-tailed fragmented DNA. Singleor dual-index adapters are ligated to the fragments, and size selection was performed with AMPureXP beads.