title: GSM1832841: 6_48 0.5µM OTA [miRNA-seq]; Danio rerio; miRNA-Seq
library_construction_protocol: Total RNA was extracted by Trizol® Reagent (Invitrogen, USA) according to the instruction manual. RNA purified was quantified at OD260nm by using a ND-1000 spectrophotometer (Nanodrop Technology, USA) and qualitated by using a Bioanalyzer 2100 (Agilent Technology, USA) with RNA 6000 labchip kit (Agilent Technologies, USA). The small RNA library construction and deep sequencing was carried out at biotechnology company (Welgene, Taipei, Taiwan). Samples were prepared using Illumina sample preparation kit according to the TruSeq Small RNA Sample Preparation Guide.The 3′ and 5′ adaptors were ligated to total RNA and reverse transcription followed by PCR amplification. The enriched cDNA constructs were size-fractionated and purified on a 6% polyacrylamide gel electrophoresis and the bands containing the 18-40 nucleotide RNA fragments (140-155 nucleotide in length with both adapters).  Libraries were sequenced on an Illumina GAIIx instrument (50 cycle single read ). Sequencing data was processed with the Illumina software