GSM5355768: Joint CIA ZT12 rep 3; Mus musculus; RNA-Seq
Joint tissue was ground with liquid nitrogen using a pestle and mortar, then transferred to a Lysing Matrix D tube containing Trizol. Tissue was homogenised using a BeadMill homogeniser (3 x 4 m/s for 40s). RNA was extracted using chloroform then precipitated using isopropanol. After washing with 75% ethanol, the RNA pellet was resuspended in RNase-free water. RNA was purified from liver samples using the SV Total RNA kit (Promega), according to the manufacturer's instructions. Tissue was homogenised using Lysing Matrix D tubes loaded into a BeadMill homogeniser (4 m/s for 20s). RNA was eluted in RNase-free water. RNA was purified from muscle samples using the ReliaPrep Tissue kit (Promega), following the manufacturer's instructions for the purification of RNA from fibrous tissue. Muscle tissue was homogenised using the same protocol as for joint, and eluted in RNase-free water. RNA libraries were prepared for sequencing using standard Illumina protocols.