Overview

This resource explains how to prepare small RNA libraries for sequencing on Illumina next-generation sequencing (NGS) systems using the Illumina miRNA Prep kit and PCR-based unique dual indexes (UDIs). The Illumina miRNA Prep kit allows for library preparation and multiplexing of up to 384 samples.

The Illumina miRNA Prep Solution facilitates both enhanced differential expression analysis using integrated unique molecular identifiers (UMIs) and discovery of novel miRNAs from cells, tissues, and biofluids. The required amount of template for a single Illumina miRNA Prep sequencing reaction can range from 1 ng to 500 ng purified total RNA.

The Illumina miRNA Prep workflow provides the following features:

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An optimized assay that does not require gel purification, excision, or elution, resulting in a reduction in hands-on time and workflow length.

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Methodology that uses modified oligonucleotides to prevent adapter dimerization in the sequencing library.

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Designed to minimize the presence of hY4 Y RNA, which is often observed in high levels in serum and plasma samples.

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Highly optimized reaction chemistry that reduces biases and background contaminants.

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Integration of unique molecular identifiers (UMIs) into the reverse transcription process enables unbiased and accurate miRNome-wide quantification of mature miRNAs by NGS systems.

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Primary and secondary data analysis solutions using the DRAGEN miRNA analysis software, developed to facilitate UMI counting, miRNA mapping, and differential expression analysis. Perform secondary analysis in the cloud via BaseSpace Sequence Hub (BSSH) and Illumina Connected Analytics (ICA).