Instrument: Illumina MiSeq
Strategy: ncRNA-Seq
Source: TRANSCRIPTOMIC
Selection: size fractionation
Layout: SINGLE
Construction protocol: Total RNA from the leaf samples was extracted using the mirVana miRNA isolation kit (Ambion, Austin, TX, USA), with modifications. Briefly, 100 mg of leaf sample was ground with 800 µl of Lysis/Binding buffer. Then, 80 µl of miRNA homogenate was added and the resulting solution was mixed by vortexing. RNA was separated by acid phenol-chloroform followed by DNase RQ1 (Promega, MA) treatment. RNA integrity was examined in a 2100 Bioanalyzer (Agilent Technologies, California, USA). For high-throughput sequencing of small RNAs, the small RNA of 15-50 nt was purified from the total RNA. The purified samples were then subjected for small RNA (sRNA) library preparation using NEBNext Multiplex Small RNA Library Prep Set for Illumina according to manufacturers instructions (New England Biolabs).