GSM7017206: T3_Transection, Triceps Brachii Muscle, Rep5; Mus musculus; RNA-Seq
Mouse skeletal muscle tissue (~30 mg) was homogenized in Trizol reagent (Invitrogen) and extracted using RNeasy mini kit (Qiagen) according to manufacturer's instructions. Total RNA concentration and integrity were determined by 260/280 and 260/230 nanodrop absorbance, as well as QUBIT and RNA6000 PicoAssay. mRNA libraries were generated using NEBNext® Ultra™ II Directional (stranded) RNA library prep kit for Illumina (NEB #E7760L) and NEBNext Poly (A) mRNA Magnetic Isolation Module (NEB #E7490). 200 ng total RNA was used for library input, fragmentation occurred for 10 minutes, and 12X PCR cycles were run. Library quality was assessed using High Sensitivity DNA Bioanalyzer and libraries were then sequenced by NovaSeq6000 SP Paired-End 150bp format at a sequencing depth of ~20 million reads per lane per sample.