GSM3591622: hSMCs_P6 Ctrl Rep2 [RNA-seq]; Homo sapiens; RNA-Seq
Total RNA was purified from hSMCs cells differentiated into myotubes (control, palmitate or TNFα treated) using AllPrep DNA/RNA/miRNA Universal Kit (Qiagen). 1 mg of total RNA was depleted of rRNA and subsequently used to generate libraries using the TruSeq standard total RNA with Ribo-Zero Gold kit (Illumina). The PCR cycle number for each library amplification was optimized by running 10% of the library DNA in a real-time PCR reaction using Brilliant III Ultra-fast SYBR Green QPCR Master Mix (AH Diagnostic) and a C1000 Thermal cycler (Bio-Rad) (Table S7). Libraries were sequencing on a Illumina NextSeq 500 system (Illumina) using the NextSeq 500/550 High Output v2 kit (75 cycles).