GSM3357362: Thirty month old_adipose_smallRNA2; Bubalus bubalis; miRNA-Seq
Total RNA was extracted using TRIzol reagent. Total RNA was assessed by electrophoresis on a denaturing agarose gel and quantified by NanoDrop spectrophotometer (NanoDrop, USA) and Agilent 2100 Bioanalyzer. The total RNA samples (3 mg) were treated with the epicentre Ribo-ZeroTM Kit to remove rRNA before constructing the RNA-seq libraries. Approximately 1 μg total RNA per sample was used for small RNA library construction according to protocol of the TruSeq® miRNA Sample Prep Kit v2 (Illumina, USA). Briefly, the 5' and 3' adaptors were ligated using T4 RNA ligase. Then the modified small RNA was subjected to RT-PCR for amplification. The 145-160 bp cDNA fragment was extracted and purified from the polyacrylamide gel electrophoresis. The enriched cDNA was sequenced in a HiSeq 2500 sequencing system (Illumina, San Diego, USA) at the Genergy Biotechnology Co., Ltd. (Shanghai, China).
GEO
GSM3357362