title: GSM9166434: myotubes, C57 vehicle sample 1, biol rep 1; Mus musculus; RNA-Seq
library_name: GSM9166434
library_construction_protocol: The myotubes (following the collection of CM) and fibroblasts (following 24 hours CM treatment) were directly lysed by adding 100 μL TRIzol (15596026, Invitrogen) for each cm2 of surface area of the culture plate. RNA was isolated using the DirectzolTM RNA miniprep kit (R2060, Zymo Research) following the manufacturer's protocol. 200ng of total RNA from each sample were sent to Novogene Co., Ltd. (Davis, CA) for sequencing.  RNA integrity was assessed using a Bioanalyzer 2100 system (Agilent Technologies). Messenger RNA was purified from total RNA using poly-T oligo-attached magnetic beads. After fragmentation, the first strand cDNA was synthesized using random hexamer primers, followed by the second strand cDNA synthesis using dTTP for non-directional library. The non-directional library, was ready after end repair, A-tailing, adapter ligation, size selection, amplification, and purification.