
Feature | Specification |
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Automation Compatible | Yes |
Product Group | Small RNA-seq |
The NEXTFLEX small RNA sequencing kit v4 uses patent-pending technology to provide a completely gel-free small RNA library preparation solution for Illumina and Element sequencing platforms.
Explore our Dharmacon™ miRNA modulation reagents to enhance or inhibit miRNA levels as part of your functional studies.
We compared different vendors, and we found that the small RNA sequencing kit from Revvity was the one that performed best.
- Dr. Sören Franzenburg, Head of NGS platform, Kiel University
The NEXTFLEX Small RNA-Seq Kit v4 offers a completely gel-free library prep workflow for Illumina® and Element® sequencers. Optimized 3′ and 5′ adapter-ligation chemistry, together with proprietary dimer-reduction oligonucleotides, minimizes adapter-dimer formation and sequence-dependent ligation bias without the need for PAGE cleanup.
The six-hour protocol reliably produces high-complexity libraries from as little as 1 ng of total RNA derived from plasma, serum, urine, CSF, cultured cells or tissue lysates. Internal benchmarking shows higher on-target mapping rates and a larger repertoire of unique miRNA species than standard small RNA workflows.
To further enhance sensitivity in biofluid studies, the kit includes tRNA and YRNA blocking oligos, reducing over-represented fragments and enriching true miRNA reads (Figures 1 & 2).
Figure 1. NEXTFLEX Small RNA-Seq Kit v4 increases miRNA reads while reducing tRNA/YRNA carry-over. Libraries from 1 ng plasma RNA were sequenced and aligned to miRNA, tRNA, and YRNA reference sets. Compared with a competitor workflow, the NEXTFLEX Small RNA kit aligns with more miRNAs while, while minimizing reads associated with t/YRNA and RNA, improving effective sequencing depth for downstream small-RNA analysis.
Figure 2: NEXTFLEX Small RNA-Seq Kit v4 detects higher number of unique miRNA species across different sample types
The gel-free NEXTFLEX Small RNA-Seq Kit v4 workflow is equally effective for exosomal RNA (exoRNA). Even when starting with the picogram-level miRNA inputs typical of exosome preparations, the protocol generates libraries with high mature-miRNA alignment rates and negligible adapter-dimer carry-over (Figure 3). This enables reliable profiling of miRNAs derived from extracellular vesicles for liquid-biopsy biomarker discovery and other low-input applications.
Internal benchmarking shows that the NEXTFLEX Small RNA-Seq Kit v4 recovers up to 45% more unique miRNA species than a competitor small RNA workflow when starting with as little as 1 ng of total RNA (Figure 4). This comprehensive profiling is essential for understanding the full spectrum of miRNA expression in each sample enabling a more complete view of post-transcriptional regulation and strengthens downstream differential-expression and biomarker-discovery analyses. The kit incorporates a highly processive rRT enzyme able to read through some of the common covalent modifications appearing in miRNA.
Figure 3. NEXTFLEX Small RNA v4 identifies 45% more unique miRNA species than competitor workflows. Venn diagram of libraries prepared from 1 ng serum RNA shows 81 miRNAs detected with both kits, an additional 53 miRNAs unique to NEXTFLEX, and 11 unique to the comparator.
The gel-free NEXTFLEX Small RNA-Seq Kit v4 readily converts ribosome-protected fragments (28–30 nt RPFs) and nascent transcripts into sequencing libraries for Ribo-Seq and nascent transcriptomics application. By adapting small RNA library preparation methods, researchers can quantify translational efficiency, map ribosome-stall sites, and reveal upstream open-reading frames, critical insights for gene-expression control, disease-mechanism studies, and drug-target discovery.
“We frequently use Revvity small RNA sequencing kits for Ribosome- and Disome-profiling. It offers a very streamlined library generation and allows for easy customization to adapt to experimental needs. With this, it consistently delivers good results. Additionally, the technical support from Revvity’s team is excellent and very helpful which further enhanced our very positive experience with this Revvity product.”
- Dr. Tobias Schmidt, CRUK Scotland Institute
Validated scripts make the NEXTFLEX workflow easy to setup on Sciclone G3 NGSx / iQ and Zephyr G3 workstation. It is also readily adapted to other liquid handlers. Up to 384 Unique Dual Index (UDI) barcodes are available, enabling high-throughput multiplexing on Illumina® and Element® sequencers while minimizing index hopping.
Boost usable miRNA reads in blood and plasma with NEXTFLEX Blood miRNA Blockers. These oligos selectively deplete highly abundant miR-486-5p, miR-92a-3p, and miR-451a—species that can consume 50–70% of reads so more sequencing depth is devoted to biologically informative miRNAs.
We have been using Revvity small RNA sequencing kits for a number of years, and we are satisfied with the results and the support received.
- Dr. Marcel van Herwijnen, Researcher/Lab Manager, Maastricht University
Pair the NEXTFLEX® Small RNA-Seq Kit v4 with Dharmacon™ miRNA mimics and inhibitors to quickly elevate or silence target miRNAs, confirm their biological impact in any cell system, and move seamlessly from expression profiling to functional validation in a single streamlined workflow.
Automation Compatible |
Yes
|
---|---|
Barcodes |
97 - 192
|
Format |
Automation Friendly Volumes
|
Product Group |
Small RNA-seq
|
Shipping Conditions |
Dual Temperature
|
Unit Size |
96 rxns
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Profiling from extracellular vesicles
Biomarker discovery/validation
Other
Ribo-Seq
Are you looking for resources, click on the resource type to explore further.
Extracellular vesicles (EVs) represent a fascinating frontier in biomarker discovery. Secreted by virtually all cell types, these...
This flyer describes the benefits of the NEXTFLEX® Small RNA-Seq Kit v4 kit
In this application note we describe a simplified, commercially available protocol encompassing exoRNA extraction and preparation...
This excel file includes the sequences of the indexes for the UDI primers included in the NEXTFLEX Small RNA-seq kit v4.
NEXTFLEX® Small RNA-Seq Kit v4 has a streamlined, gel-free workflow delivering exceptional miRNA mapping and discovery rates even...
This poster illustrates the effieciency of the NEXTFLEX® Small RNA-Seq Kit v4 kit of discovering biomarkers.
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