Instrument: Illumina HiSeq 2500
Strategy: miRNA-Seq
Source: TRANSCRIPTOMIC
Selection: size fractionation
Layout: SINGLE
Construction protocol: Total RNA was respectively extracted from frozen pectoral muscle samples with TRIzol reagent (Invitrogen, USA) according to the manufacturer's protocol. The quantity of total RNA was assessed using an Agilent 2100 Bioanalyzer with Agilent RNA 6000 nano Reagents Part 1 kit (Agilent Technologies, USA). Each development stage has three replicates that came from the three pigeons. Total RNA of each replicate was individually used for library construction. 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, libraries were sequenced by an Illumina Hiseq 2500 platform and generate 50 bp single-end reads.