title: GSM8775548: Male Muscle Rapamycin Biol Rep 4; Mus musculus; RNA-Seq
library_construction_protocol: Total RNA was isolated from 4-10mg of spleen, muscle and kidney using TrIzol. Samples were treated with DNAase using the Turbo DNAse kit (ThermoFisher Scientific) to remove genomic DNA. poly(A)+ mRNA was isolated using the NEBNext Poly(A) mRNA Magnetic Isolation kit (New England BioLabs, Prior to first strand synthesis, samples were randomly primed (5' d(N6) 3' [N=A,C,G,T]) and fragmented. First strand synthesis was done using the Protoscript II Reverse Transcriptase for approximately 40 minutes at 42⁰C. All remaining steps for library construction were done according to the NEBNext Ultra™ II Non- Directional RNA Library Prep Kit for Illumina, RNA Seq with 50 million 150 bp paired-end reads/sample