Instrument: NextSeq 500
Strategy: miRNA-Seq
Source: TRANSCRIPTOMIC
Selection: size fractionation
Layout: SINGLE
Construction protocol: The obtained liver tissues were minced quickly, mixed at a ratio of 1:10 with QIAzol lysis reagent and homogenized by bead beating. Total RNA was extracted according to the manufacturer's instruction of the QIAzol lysis reagent with the exception that chloroform was replaced by 1-bromo-3-chloropropane for the phase separation step. To remove genomic DNA contamination in RNA samples, an enzymatic digestion step using DNase I was carried out. Finally, total RNA was re-suspended in RNase-free water. RNA quantity and RNA quality number (RQN ≥ 8) was determined by the Fragment Analyzer CE12. Five libraries were generated from five mice (three mice from E. multilocularis-infected group and two mice from the uninfected control group). The total RNA was checked on an Agilent 2100 Bioanalyzer instrument for degradation. Subsequently, the CleanTag Ligation Kit (was used to prepare small RNA stranded libraries from total RNA (1µg RNA per library). Libraries were analyzed a second time on the Bioanalyzer to check for the expected miRNAs fragment peak at 141 bp and subsequently a Sage Science Pippin Prep instrument was used to select the expected fragment size range. In a last step before the deep sequencing, the quality and concentration of each final library (the eluted 141 bp band) were submitted to PicoGreen analysis.