title: miRNA-Seq of Humulus lupulus (Wye Target):control, biol. rep. 2
design: Small RNA libraries were constructed using the Ion Total RNA-Seq Kit v2 and Ion Xpress RNA-Seq Barcode 1-16 Kit following the manufacturers instructions. Briefly, adaptors were hybridized and ligated to small RNAs, and the reverse transcription was performed. Afterwards, purification and size-selection were performed using magnetic beads to obtain only miRNAs and other small RNAs to which barcodes were added through PCR amplification. The yield and size distribution of amplified cDNA libraries were assessed with Agilent 2100 Bioanalyzer instrument (Agilent Technologies, Inc., Santa Clara, California, USA) and Agilent High Sensitivity DNA Kit to pool equimolar barcoded libraries of each cultivar separately. Three inoculated and three mock-inoculated barcoded libraries of susceptible or resistant cultivars were pooled in equimolar concentration and prepared for sequencing according to the manufacturers instructions, accompanying Ion PI HiQ OT2 200 Kit and Ion PI Hi-Q Sequencing 200 Kit. Both prepared samples were sequenced on the Ion Proton System (Waltham, Massachusetts, USA).