title: Identification and characterization of the Chinese giant salamander (Andrias davidianus) miRNAs by deep sequencing and their targets predication
design: Total RNA was extracted from animal tissues using TRIzol reagent (Invitrogen, USA) and then was treated with RNase-free DNase I (Takara, China) to remove any contaminating genomic DNA according to the manufacturer's protocol. The quantity and purity of total RNA was monitored via analysis by NanoDrop ND-1000 spectrophotometer (Nano Drop, USA) at 260/280 nm (ratio=2.0). The integrity of total RNA was tested via analysis by Bioanalyzer 2100 and RNA 6000Nano LabChip Kit (Agilent, USA) with RIN number>7.0. Briefly, small RNAs of 18-35 nt in length were first isolated from the total RNA by size fractionation. Then, these small RNAs were ligated with 5'-RNA and 3'-RNA adapters and subsequently reverse transcription PCR was used to create cDNAs. The amplified cDNAs were purified and sequenced with Illumina Hiseq 2500 platform.