title: The third longissimus dorsi muscle tissue sample of 1-month-old Hanper sheep.
design: In this experiment, RNA was extracted from longissimus dorsi muscle tissue using the Trizol reagent Kit (Invitrogen, Carlsbad, CA, USA) following the manufacturer's protocol. The RNA integrity was assessed using an Agilent 2100 Bioanalyzer (Agilent Technologies, Palo Alto, CA, USA), and RNA samples meeting the quality criteria (0.5 g) were used for miRNA library construction. Small RNA fragments (18-30 nt) were separated and recovered using PAGE technology. After adapter ligation to both ends of the small RNA, miRNAs were enriched by reverse transcription PCR. The bands of approximately 140 bp were recovered, purified, and submitted for library preparation. Library quality control was performed, and sequencing was carried out on the Illumina platform (Beijing NoVo gene Bioinformatics Technology Co., Ltd.). RNA degradation and contamination were checked via 1% agarose gel electrophoresis, while RNA purity was assessed using a NanoPhotometer spectrophotometer (IMPLEN, CA, USA). RNA concentration was measured with the Qubit RNA Assay Kit on a Qubit 2.0 Fluorometer (Life Technologies, CA, USA). RNA integrity was further evaluated using the RNA Nano 6000 Assay Kit on the Agilent Bioanalyzer 2100 system. For small RNA library construction, 3 g of total RNA per sample was used as the starting material. Libraries were prepared using the NEBNext Multiplex Small RNA Library Prep Set for Illumina (NEB, USA), with index sequences added for sample identification. The quality of the libraries was assessed on the Agilent Bioanalyzer 2100 using High Sensitivity DNA Chips. Clustering of the index-coded samples was performed on a cBot Cluster Generation System using the TruSeq SR Cluster Kit v3-cBot-HS (Illumina), following the manufacturer's instructions. Sequencing was carried out on the Illumina HiSeq 2500 platform, generating 52 bp single-end reads.
identifiers.namespace: SUB15166192
identifiers.id: M1_3