miRNA-Seq of macrophage-derived exsomes in spinal tuberculosis and disc degeneration individuals bone marrow blood
Total RNA was extracted from exosomes using TRIzol reagent (Invitrogen, cat. NO 15596026) following the methods described by Chomczynski et al [12]. After RNA extraction, DNA digestion was performed using DNaseI. RNA quality was assessed by determining the A260/A280 ratio using a NanodropTM OneC spectrophotometer (Thermo Fisher Scientific Inc). Additionally, RNA integrity was confirmed by 1.5% agarose gel electrophoresis. The qualified RNAs were quantified using Qubit3.0 and the QubitTM RNA Broad Range Assay Kit (Life Technologies, Q10210). Total RNA (3 L) was used as input for miRNA library preparation using the KC-DigitalTM small RNA Library Prep Kit for Illumina (Catalog No. DR08602, Wuhan Seqhealth Co., Ltd. China) according to the manufacturer's instructions.