An Improved Sample-to-Sequence Workflow for Fast and Robust miRNA Analysis on the Ion Torrent PGM Using the TaqMan? miRNA ABC Purification Kit

摘要

MicroRNA (miRNA) profiling via next generation sequencing is a promising new genome-wide approach for quantifying miRNAs with single nucleotide resolution, making it an attractive technology for diagnostic applications. However, sample processing and miRNA extraction methods can have a significant impact on miRNA expression profiling, particularly for samples that are prone to degradation. Moreover, current miRNA purification methods are tedious, time-consuming, and susceptible to variability, which can lead to poor performance in downstream steps. Here we demonstrate that the TaqMan? miRNA ABC Purification Kit can be used in combination with the Ion Torrent Total RNA-Seq v2 kit to provide an integrated sample-to-sequence workflow that also offers improved assay performance. With its unique bead-capturing technology, the TaqMan? miRNA ABC Purification kit dramatically simplifies and improves the robustness of miRNA isolation, especially from difficult sample types without compromising critical assay parameters such as accuracy, sensitivity, and reproducibility. In addition, the kit eliminates the small RNA enrichment step from the Total RNA-Seq kit v2 workflow, which completely obviates the need for gel size selection during small RNA library preparation.