Abstract
The small RNA (<50 nts) transcriptome contains a diverse array of RNA molecules containing a variety of different 5′ end modifications such as 5′-monophosphate (5′ pN), 5′- triphosphate (5′ pppN), or 5′-cap (GpppN). Such 5′ end diversity makes it difficult to capture the entire small RNA transcriptome using conventional small RNA-seq library preparation methods. In addition conventional methods generate a significant amount of adaptor-dimer that contaminates the library. We demonstrate a greatly improved small RNA–Seq library preparation process, the ScriptMiner™ technology, that enables capture of the entire small RNA transcriptome. Additionally, the ScriptMiner procedure includes novel chemistry for greatly reducing the amount of undesired adaptor-dimers in the final library and ScriptMiner libraries are readily barcoded with Illumina® Index reads or user-defined barcodes. The result is a more sensitive and comprehensive representation of the small RNA transcriptome in the cell.