GSM3665327: SMAdel7_HetP6_R1; Mus musculus; RNA-Seq
RNA was isolated from whole heterozygote control or homozygote SMN7 mouse spinal cords (P2, P6, or P10) by Trizol/chloroform extraction. Tissues were dissociated in 1mL Trizol using a bead beater homogenizer (Precellys) using 1.4 mm Zirconium oxide beads (5500–3x-20s-030). Homogenized samples were then incubated for 5 minutes at room temperature (RT) followed by addition of 200uL chloroform and vigorous agitation for 15 seconds. Samples were then incubated for an addition 3 minutes at RT then centrifuged at 12,000 rcf for 15 minutes at 4C, after which the aqueous layer was collected for subsequent clean-up using an RNeasy isolation kit (Qiagen) with DNase treatment according to manufacturer's instructions with extraction in 50 μl RNAse-free water. RNA concentration was measured by nanodrop and submitted for sequencing. RNA libraries were prepared using the TruSeq Stranded Total RNA Library Prep Kit with RiboZero (Illumina)