This whitepaper shows that small RNA sequencing libraries can be prepared successfully from plant total RNA from a variety of species using the NEXTflex® Small RNA Seq Kit v3 gel-free protocol.  Ligation of a 3’ adapter via the 3’ OH of the miRNA molecule is the first step in small RNA NGS library construction. However, plant miRNAs have a 2’-O-methylation which reduces ligation efficiency of the 3’ OH, making plant miRNA libraries difficult to generate. Here, we show that miRNA profiling from agricultural crops using total RNA inputs can be successfully achieved using the NEXTflex Small RNA Sequencing Kit v3, without a PAGE gel purification step, and that extending the ligation incubation step increases library yield.

To learn more about this bias-reducing technology and how it can be used to increase the efficiency and discovery of miRNAs for small RNA NGS sequencing library prep with plant samples, read our recent whitepaper.

For research use only. Not for use in diagnostic procedures.