Transcriptome analysis of differentially expressed miRNAs in bovine skeletal muscle
Total RNA was extracted from ovary tissue samples by using Trizol reagent (TaKaRa, Dalian, China). Gel electrophoresis, Agilent 2100 and NanoDrop methods were used to detect integrity and concentration of RNA samples. Before constructing the RNA-seq libraries, the epicenter Ribo-ZeroTM Kit (Illumina, San Diego, CA, USA) was used to remove rRNA. Briefly, total RNA was purified by polyacryl-amide gel electrophoresis (PAGE) to enrich the sRNAs with lengths of 1535 nt, then the sRNAs were ligated with adapters and amplified by RT-PCR. The amplification products were then separated on a PAGE gel, and the transcriptome sequencing was performed on the HiSeq 2000 (Illumina, California, USA).