GSM3895742: articular cartilage, WT rep3; Mus musculus; RNA-Seq
Cartilage was stored at -20oC in Allprotect Tissue Reagent (Qiagen, Valencia, CA) immediately after resection from the subchondral bone. For RNA isolation, cartilage was pulverized in a 6770 Freezer/Mill Cryogenic Grinder (SPEX SamplePrep, Metuchen, NJ), and homogenized in Qiazol Lysis Reagent (Qiagen) using 25mg tissue per 700ul Qiazol. RNA was isolated using the miRNeasy Mini kit (Qiagen) with on-column DNAse digestion, followed by removal of proteoglycans using RNAmate (BioChain Institute, Newark, CA). 5Mouse articular cartilage was collected from both sides of the femoral condyle and tibial plateau from 3-month-old mice. The cartilage was homogenized and incubated in TRIzol (Invitrogen) at room temperature. After addition of chloroform, samples were centrifuged, and the supernatant was collected for RNA isolation. Libraries were prepared using the SMART-Seq v4 Ultra Low Input RNA Kit (Clontech).