GSM5144864: osteoarthritic cartilage, s7; Homo sapiens; RNA-Seq
For RNA isolation, about 1g of cartilage was pulverized in liquid nitrogen and homogenized in Trizol at a concentration of 1g tissue per 10ml Trizol (Invitrogen), followed by incubation at 4oC with rotating for 2h. To remove debris, samples were mixed with 0.2 volumes of chloroform, vortexed for 20sec, and centrifuged at 14000 rpm for 20min at 4 oC. The aqueous phase was collected and gently mixed with an equal volume of isopropanol, followed by centrifugation at 14000 rpm for 20min at 4 oC. The resulting pellet was suspended in 350µl of RLT buffer and processed for cleanup using the RNeasy Mini Kit (Qiagen) in accordance with manufacturer's instructions. RNA libraries were prepared for sequencing using standard Illumina protocols