GSM5398369: Flight-F03-qM20; Mus musculus; RNA-Seq
Frozen carcasses were thawed on wet ice before tissue dissection. Skeletal muscles from the left hind limb of each carcass were individually dissected and preserved in RNAlater preservative solution. Total RNA was isolated using the acid guanidinium thiocyanate-phenol-chloroform extraction followed by silica-membrane purification. Briefly, frozen tissue samples were minced into small pieces (1mm x 1mm x 1mm). A homogeneous lysate was achieved by adding lysing buffer and gentle ultrasound vibration on ice. Tissue lysate was centrifuged and the supernatant was used for RNA phenol/chloroform extraction. After phase separation, the aqueous layer was transferred and mixed with an equal volume of 70% ethanol. Total RNA was then extracted using RNeasy spin columns from the RNeasy Micro Kit (Qiagen, Hilden, Germany) according to the manufacturer's protocol. RNA libraries were prepared for sequencing using standard Illumina protocols