GSM4396419: SMSC-1d; Columba livia; miRNA-Seq
The total RNA was harvested using Trizol-LS reagent. The quantity of total RNA was assessed using an Agilent 2100 Bioanalyzer with Agilent RNA 6000 nano Reagents Part 1 kit (Agilent Technologies, USA). For samples of high-throughput sequencing, the RNAs from three replicates at each differentiation stage were pooled as one RNA sample. For each library, small RNA ranging from 10-45 nt in length was purified by polyacrylamide gel electrophoresis and ligated using proprietary adaptors. The modified small RNA was then reverse-transcribed to cDNA and amplified by PCR. Finally, the libraries were sequenced on a BGIseq-500 sequencing platform.