GSM7789928: 32_RORA CTRL_ZT20; Mus musculus; RNA-Seq
Total RNA from muscle tissues was extracted using a hybrid method combining TRI-Reagent (Sigma-Aldrich T9424) and RNeasy Mini Kit (QIAGEN 74104). RNA quality was determined on the Bioanalyzer instrument (Agilent Technologies, Santa Clara, CA, USA) using the RNA 6000 Nano Chip (Agilent, Cat# 5067-1511) and quantified by Spectrophotometry using the NanoDrop ND-1000 Instrument (NanoDrop Technologies, Wilmington, DE, USA). Library preparation was performed with 1μg total RNA using the TruSeq Stranded mRNA Library Prep Kit High Throughput (Cat# RS-122-2103, Illumina, San Diego, CA, USA). Libraries were quality-checked on the Fragment Analyzer (Advanced Analytical, Ames, IA, USA) using the Standard Sensitivity NGS Fragment Analysis Kit (Cat# DNF-473, Advanced Analytical) revealing excellent quality of libraries (average concentration was 152±9 nmol/L and average library size was 374±4 base pairs). Samples were pooled to equal molarity. Each pool was quantified by PicoGreen Fluorometric measurement in order to be adjusted to 1.8pM and used for clustering on the NextSeq 500 instrument (Illumina).