GSM7506195: N2.IP; Homo sapiens; OTHER
Knee cartilage samples of medial condyles were collected from three patients who underwent knee arthroplasty due to advanced OA and three patients who underwent thigh amputation due to trauma. Fresh samples were immediately frozen in liquid nitrogen and stored at -80 ℃ for detection. The total RNA from each sample was extracted using TRIzol reagent (Invitrogen Corporation, Carlsbad, CA). The content of rRNA in the total RNA was reduced using the Ribo-Zero rRNA Removal Kit (Illumina, Inc., CA, USA). The concentration of total RNA was measured using a NanoDrop ND-1000 spectrophotometer (NanoDrop Technologies, Wilmington, DE, USA). RNA with an OD260/OD280 ratio range of 1.8–2.1 was marked as acceptable. The m5C-IP-Seq service was provided by CloudSeq Inc. (Shanghai, China). Total RNA was subjected to immunoprecipitation with the GenSeq® m5C-IP Kit (GenSeq Inc.) by following the manufacturer's instructions. RNA libraries for IP and input samples were then constructed with the GenSeq® Low Input Whole RNA Library Prep Kit (GenSeq, Inc.) by following the manufacturer's instructions. Libraries were qualified using the Agilent 2100 bioanalyzer and then sequenced on a NovaSeq platform.. MeRIP-seq