NC 6
Solid tissue samples were either immediately stabilized in RNAlater (Qiagen, Germany) and then stored at 70C or fixed in formalin and embedded in paraffin blocks. RNA extraction was performed immediately before the preparation of sequencing libraries using QIAGEN RNeasy Kit (Qiagen) or Direct-zol RNA MiniPrep (Zymo Research) with TRI Reagent (MRC) for tissues in RNAlater and the RecoverAll RNeasy FFPE Kit (Qiagen), following the manufacturer's protocol. RNA Integrity Number (RIN) was measured using Agilent 2100 bioanalyzer. Agilent RNA 6000 Nano or Qubit RNA Assay Kits were used to measure RNA concentration. KAPA RNA Hyper with RiboErase (KAPA Biosystem) Kit was used for further depletion of ribosomal RNA and library preparation. Different adaptors were used for multiplexing samples in one sequencing run. Library concentrations and quality were measured using Qubit ds DNA HS Assay kit (Life Technologies) and Agilent Tapestation (Agilent).