Search all

Revvity Sites Globally

Select your location.

*e-commerce not available for this region.

Australia

Austria

Belgium

Brazil *

Canada

China *

Denmark

Finland

France

Germany

Hong Kong *

India *

Ireland

Italy

Japan *

Luxembourg

Mexico *

Netherlands

Norway

Philippines *

Republic of Korea *

Singapore *

Spain

Sweden

Switzerland

Thailand *

United Kingdom

United States

NEXTFLEX Small RNA Sequencing Kit V4

Name: NEXTFLEX Small RNA Sequencing Kit V4
Price: 7198.52 USD
Availability: InStock

SDS

IFUs, Manuals & more

The NEXTFLEX® Small RNA-Seq Kit v4 enables labs to reduce bias in their Illumina® and Element Biosciences® small RNA sequencing workflows while leveraging a completely gel-free protocol, even with low input samples.

View product information View Related reagents

Feature	Specification
Automation Compatible	Yes
Product Group	Small RNA-seq

Request More Information

View Related reagents

View product information View Related reagents

Product Variants

Unit Size: 8 rxns

Barcodes

1 - 8

Part #:

NOVA-5132-31

List Price

USD 781.09

Your online price:

Unit Size: 8 rxns

Barcodes

9 - 16

Part #:

NOVA-5132-31C

List Price

USD 781.09

Your online price:

Unit Size: 48 rxns

Barcodes

1 - 48

Part #:

NOVA-5132-32

List Price

USD 3,904.56

Your online price:

Unit Size: 96 rxns

Barcodes

1 - 96

Part #:

NOVA-5132-41

List Price

USD 7,198.52

Your online price:

Unit Size: 96 rxns

Barcodes

97 - 192

Part #:

NOVA-5132-42

List Price

USD 7,198.52

Your online price:

Unit Size: 96 rxns

Barcodes

193 - 288

Part #:

NOVA-5132-43

List Price

USD 7,198.52

Your online price:

Unit Size: 96 rxns

Barcodes

289 - 384

Part #:

NOVA-5132-44

List Price

USD 7,198.52

Your online price:

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

Total list price:

USD 7,198.52

Supporting products you might need

Are you working with blood samples?

Revvity kit box for reagents and consumables

Add to bundle View details

NEXTFLEX blood miRNA blockers

Part #: NOVA-513103

List Price: USD 453.68

Your online price:Login to view

Product information

Overview

The NEXTFLEX small RNA sequencing kit v4 uses patent-pending technology to provide a completely gel-free small RNA library preparation solution for Illumina sequencing platforms.

Completely gel-free protocol from purified miRNA or 1 ng of total RNA
Automated on the Sciclone® G3 NGSx, Sciclone G3 NGSx iQ™, and the Zephyr® G3 NGS workstations
Patent pending technology optimized for challenging samples such as plasma or serum, including exosomes
Exceptional miRNA discovery
Ready to sequence libraries in ~6 hours
Multiplexing of up to 384 samples
An automation protocol is available
Functionally certified with Illumina sequencing platforms

Explore our Dharmacon^™ miRNA modulation reagents to enhance or inhibit miRNA levels as part of your functional studies.

Additional product information

Robustness across diverse sample types, from tissues to biofluids
Streamlined workflow without gel cuts
Fully automated - easy scalability from small to large studies
Protocols available to select for small RNA species other than miRNA

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

Gel-free small RNA library prep with reduced bias

The NEXTFLEX small RNA sequencing kit v4 uses patent-pending technology to provide a completely gel-free small RNA library preparation solution for Illumina sequencing platforms. Our approach to reducing bias involves optimization of the ligation reaction and the use of dimer reduction oligonucleotides. These changes result in decreased bias in comparison to standard protocols.

NEXTFLEX Small RNA-Seq Kit v4 allows robust preparation of libraries from diverse sample types in approximately 6 hours, with as little input as 1 ng of total RNA. Libraries prepared with this kit have a higher proportion of mapping reads and unique miRNAs than leading competitors. The kit contains tRNA and YRNA blockers to efficiently remove these species in biofluid samples (Figure 1 and 2).

Figure 1: Gel-free miRNA library prep from various biofluids

Figure 2: Gel-free miRNA library prep from exosomes

This convenient gel-free workflow incorporated in the NEXTFLEX Small RNA-Seq Kit v4 enables the construction of libraries from exoRNA. Using this workflow, researchers can achieve a high number of reads aligning to mature miRNA and low adapter dimers, even with the very low miRNA inputs characteristic of exosome samples (Figure 3).

Figure 3: Average top 10 miRNA observed with exosomes from bone marrow and pooled serum samples.

Improved miRNA discovery

The performance of small RNA sequencing is heavily dependent on the workflow used. The NEXTFLEX® Small RNA Sequencing Kit v4 can identify a significantly higher number of unique miRNA species compared to other available solutions, even at low inputs (Figure 4). This comprehensive profiling is essential for understanding the full spectrum of miRNA expression in each sample. Detecting even the low-expressed miRNA present in the sample provides deeper insights into regulatory mechanisms and facilitates biomarker discovery.

Serum-Kit-Comparison-Venn-Diagram-Larger-Numbers

Figure 4. Venn diagram showing that 45% more unique miRNA species were identified when using the NEXTFLEX^® Small RNA-Seq Kit v4 than Competitor A kit. Input was 1 ng of RNA extracted from serum.

Ribosome Profiling (Ribo-Seq) Application

The NEXTFLEX Small RNA-Seq v4 Kit can also be used for ribosome profiling (Ribo-Seq), which analyzes ribosome-protected mRNA fragments to study translation. By adapting small RNA library preparation methods, researchers can gain insight into gene regulation and translation dynamics, making Ribo-Seq valuable in fields like gene expression, disease mechanisms, and drug 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

Small RNA sequencing library prep optimized for automation

By eliminating all gel-based steps even for low input samples, the NEXTFLEX Small RNA-Seq Kit V4 is optimized for easy migration onto automated liquid handling platforms. This workflow is automated on the Sciclone G3 NGSx, Sciclone G3 NGSx iQ, and the Zephyr G3 NGS workstations

Illumina® and Element® small RNA sequencing multiplexing with UDI barcodes

Up to 384 UDI barcoded primers are included in each kit for multiplexing. These UDIs allow libraries to be run on any Illumina and Element Biosciences sequencing platform. These include the Illumina NextSeq® and Novaseq® sequencers and only require standard Illumina sequencing primers.

Maximize miRNA mapped reads with NEXTFLEX blood miRNA blockers

The NEXTFLEX blood miRNA blockers deplete common miRNA in blood and plasma such as miR-486-5p, miR-92a-3p and miR-451a. These constitute 50-70% of the small RNA present in those samples and usually are not of interest for researchers.

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

Listen to the Experts:

Listen to our webinar, "Exploring Circulating Extracellular Vesicles in Liquid Biopsies for Neurological Disorders," where Bessie Meechoovet, a Senior Research Associate from the Jensen lab at the Translational Genomics Research Institute (TGen), explains how cell-free biomarkers are used to enhance our understanding of neurodegenerative diseases.

The Jensen lab utilizes long-read single-cell and RNA transcriptomics data to identify minimally invasive biomarkers in biofluids. Ms. Meechoovet describes a method they've developed to enrich brain extracellular vesicles, aiding in the discovery of potential biomarkers.

Specifications

Other Specifications

Automation Compatible	Yes
Barcodes	289 - 384
Format	Automation Friendly Volumes
Product Group	Small RNA-seq
Shipping Conditions	Dual Temperature
Unit Size	96 rxns

Citations

Almeida, M.V., Blumer, M., Yuan, C.U. et al. Dynamic co-evolution of transposable elements and the piRNA pathway in African cichlid fishes. Genome Biol 26, 14 (2025). https://doi.org/10.1186/s13059-025-03475-z.
Apostolov, A., Mladenović, D., Tilk, K. (2025) Multi-omics analysis of uterine fluid extracellular vesicles reveals a resemblance with endometrial tissue across the menstrual cycle: biological and translational insights, Human Reproduction Open, https://doi.org/10.1093/hropen/hoaf010.
Chu, C. P., Nabity, M. B. (2025) Technical considerations and review of urinary microRNAs as biomarkers for chronic kidney disease in dogs and cats. Vet Clin Pathol. 2025; 00: 1-19. doi:10.1111/vcp.13412.
Karere, G. M., Hsu, F.-C. Hepple, R. T. et al. (2025) MicroRNA signatures of VO2peak in older adult participants of the Study of Muscle, Mobility and Aging. bioRxiv 2025.01.08.631999; doi: https://doi.org/10.1101/2025.01.08.631999.
Lill C. M., et al. (2025) EPIC4ND: European Prospective Investigation into Cancer and Nutrition follow-up for neurodegenerative diseases. medRxiv 2025.01.29.25321340; doi: https://doi.org/10.1101/2025.01.29.25321340.
Marocco. F. (2025) The RNA binding protein PCBP2 is a regulator of miRNAs partition between cell and extracellular vesicles. [Doctoral dissertation, Sapienza University of Rome]. IRIS Uniroma1. https://iris.uniroma1.it/retrieve/6d5c8022-59e2-44ec-b8b6-eb7a6ea8cc6e/….
Rossner, p., Javorkova, E., Sima, M. et al. Skin wound healing: the impact of treatment with antimicrobial nanoparticles and mesenchymal stem cells. (2025) Authorea. February 17, 2025. DOI: 10.22541/au.173977945.54128573/v1.
Tahiri, I., Llana, S.R., Díaz-Castro, F. et al. AgRP neurons shape the sperm small RNA payload. Sci Rep 15, 7206 (2025). https://doi.org/10.1038/s41598-025-91391-4.

FAQs

What types of RNAs are included in the Small RNA library?

The NEXTFLEX Small RNA Sequencing kit v4 is designed to capture short RNAs (17-65 nucleotides) with 5' monophosphorylated and 3' hydroxylated ends. The kit also allows for the capture of larger molecules (<200 bp) by using the no size selection protocol. MicroRNAs typically have a 5' monophosphate group after cleavage by the enzyme Dicer. Other molecules, such as tRNA and yRNA, also have a 5' monophosphate at their end and they will be incorporated in the libraries if present in the sample, such as serum and plasma. The kit includes blocking oligos to prevent unwanted tRNAs and yRNA from being incorporated into the library. For removal of other types of RNA, please contact our technical support team.
Do you have any recommendations for extraction of RNA for Small RNA sequencing?

Any extraction method that generates a high-purity RNA preparation while preserving the fraction of RNA molecules with size <200 nt is compatible with small RNA sequencing. Popular extractions method used together with our NEXTFLEX® Small RNA Sequencing kit v4 include, but are not limited to, chemagic™ miRNA kits (Revvity), miRNeasy Kits (QIAGEN), MagMAX™ mirVana™ (Thermofisher) and MicroRNA Purification kit (Norgen Biotek).
Are the libraries produced by NEXTFLEX Small RNA Sequencing Kit directional?

Yes, the NEXTFLEX Small RNA libraries are directional. During the library preparation 5' and 3' adapters are ligated to the RNA molecule. This ensures that the resulting cDNA retains the original RNA strand's orientation.
Can I use total RNA, or do I have to isolate or enrich the sample for small RNA?

Small RNA libraries can be prepared using total RNA. Enriched small RNA samples can be used but are not necessary. Total RNA with a RIN (RNA Integrity Number) higher than 7 is recommended.
What happens if the quality of my RNA is very low?

The NEXTFLEX Small RNA v4 is a gel-free workflow designed to remove adapter dimers (libraries with no insert). However, if the input sample is heavily degraded, it will contain a significant proportion of fragments with size <16 nt. Those fragments will appear in the final library preparation and will decrease the percentage of reads mapping small RNA. To improve mapping rate there are two options: one option is to perform gel electrophoresis to separate the band of intact small RNA from other fragments, followed by gel band excision and purification. The second option is to use an upstream kit to remove degraded RNAs before entering library preparation. Please contact Revvity technical support team to discuss if this is needed and the best approach for your project.
How should my NEXTFLEX Small RNA libraries be trimmed?

Please download the trimming instructions provided.
What should I do if my Small RNA may not have 5' Phosphate and 3' OH groups?

Please contact Revvity technical support for recommendations on treatment of the RNA using T4 Polynucleotide Kinase (T4 PNK).
How are the barcodes introduced in the NEXTFLEX Small RNA libraries?

Two unique 8 bp indices are introduced during the PCR step (Step F).
What are the recommended settings for sequencing?

Due to the size of small RNAs, a read of 50 bp is sufficient to cover the entire insert and includes enough of the adapter to be identified and trimmed during data analysis. Single or paired-ended is possible, although paired-end generally does not add additional information. Care should be taken to ensure the number of cycles does not exceed the total length of the library molecule (insert + adapter). This will cause low quality sequencing cycles which can result in lower quality data or no data generation.
What is the number of reads required for small RNA sequencing?

The number of reads required for small RNA sequencing typically ranges from 1 to 5 million reads per sample. This range can vary depending on the specific goals of your experiment, for example differential gene expression or microRNA discovery typically require greater numbers of reads. In addition, the complexity of the input tissue and amount of starting microRNAs (or other target RNAs) may require higher number of sequencing reads. If in doubt, please contact Revvity technical support team for additional information on your particular project.
Do I have to use custom Illumina® sequencing primers for the libraries generated with NEXTFLEX Small RNA Sequencing Kit v4?

No. NEXTFLEX Small RNA v4 libraries are compatible with Illumina sequencing reagents, therefore designing custom sequencing primers is not necessary.
What do I need to do to run the NEXTFLEX Small RNA libraries on the AVITI™ sequencer from Element® Biosciences?

NEXTFLEX Small RNA v4 libraries can run native on Element Cloudbreak™ Freestyle Sequencing Kits, without any extra step.
Are the Monarch™ Spin RNA Cleanup Kits (NEB) compatible with NEXTFLEX reagents for RNA library prep?

Yes. RNA purified with this kit can be used for small RNA library preparation using NEXTFLEX Small RNA Sequencing Kit v4. This kit has a 15 nt cut off, which means that eluates from this kit will increase significantly the proportion of mapped reads for small RNA and is a convenient option to improve the percentage of small RNA mapped reads in very degraded samples.
Do you have recommendations for analysis of small RNA sequencing data?

General recommendations for analysis of small RNA data are provided in our blog "Navigating the complexities of miRNA sequencing data analysis". For specific questions, or if you are new to this field, please contact our technical support team.

Resources

Are you looking for resources, click on the resource type to explore further.

1-5 of 5 Resources

Flyer

Gel-free small RNA library prep kit for Illumina sequencing

This flyer describes the benefits of the NEXTFLEX® Small RNA-Seq Kit v4 kit

Learn More

Application Note

miRNA Exosomes App Note

In this application note we describe a simplified, commercially available protocol encompassing exoRNA extraction and preparation...

Learn More

Application Note

Simplify Low-Input Small RNA-seq Library Prep and Improve Discovery Application Note

NEXTFLEX® Small RNA-Seq Kit v4 has a streamlined, gel-free workflow delivering exceptional miRNA mapping and discovery rates even...

Learn More

Scientific Poster

Small RNA Sequencing and Biomarker Discovery

This poster illustrates the effieciency of the NEXTFLEX® Small RNA-Seq Kit v4 kit of discovering biomarkers.

Learn More

Application Note

Unlocking the potential of small RNA biomarkers with Revvity’s automated workflow

In the rapidly evolving field of molecular biology, the extraction and analysis of small RNA from plasma have become pivotal for...

Learn More

SDS, COAs, Manuals and more

Are you looking for technical documents related to the product? We have categorized them in dedicated sections below. Explore now.

Safety data sheet

Language
English
Country
United States
Name
MSDS 5132 NEXTflex Small RNA V4.pdf

IFUs, Manuals & more

Resource Type
Barcode Indexes
Language
English
Country
-
Name
NEXTFLEX UDI primers small RNA seq kit v4
Resource Type
Manual
Language
English
Country
-
Name
NEXTFLEX® Small RNA-Seq Kit V4 user Manual
Resource Type
Manual
Language
English
Country
-
Name
NEXTFLEX Small RNA v4 Trimming Instructions
Resource Type
Manual
Language
English
Country
-
Name
NEXTFLEX Small RNA v4 Supplementary No Size Selection Protocol

Related reagents

0 of 2 selected

miRNA Extraction Kits

CMG-1223

chemagic miRNA 1K Kit H24

Part number:CMG-1223

List price:USD 5,804.34/each

Your online price:Log in to view

Total list price:

USD 0

CMG-1224

chemagic miRNA 200 Kit H96

Part number:CMG-1224

List price:USD 9,708.43/each

Your online price:Log in to view

Total list price:

USD 0

0 of 2 selected

How can we help you?

We are here to answer your questions.

Contact sales I would like more product information Contact customer care Get technical support

Revvity Sites Globally

About the product

Specifications

About the product

USD 7,198.52

NEXTFLEX blood miRNA blockers

Product information

Overview

Additional product information

We compared different vendors, and we found that the small RNA sequencing kit from Revvity was the one that performed best.

Gel-free small RNA library prep with reduced bias

Improved miRNA discovery

Ribosome Profiling (Ribo-Seq) Application

Small RNA sequencing library prep optimized for automation

Illumina® and Element® small RNA sequencing multiplexing with UDI barcodes

Maximize miRNA mapped reads with NEXTFLEX blood miRNA blockers

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.

Listen to the Experts:

Specifications

Citations

SDS, COAs, Manuals and more

Related reagents

miRNA Extraction Kits

Explore BioLegend

March lab savings - SAVE 10%

March lab savings - SAVE 10%

March lab savings - SAVE 10%

March lab savings - SAVE 10%

March lab savings - SAVE 10%

March lab savings - SAVE 10%

March lab savings - SAVE 10%

March lab savings - SAVE 10%

March lab savings - SAVE 10%

March lab savings - SAVE 10%

March lab savings - SAVE 10%

March lab savings - SAVE 10%

March lab savings - SAVE 10%

March lab savings - SAVE 10%

March lab savings - SAVE 10%

March lab savings - SAVE 10%

March lab savings - SAVE 10%

March lab savings - SAVE 10%

March lab savings - SAVE 10%

March lab savings - SAVE 10%

March lab savings - SAVE 10%

March lab savings - SAVE 10%

March lab savings - SAVE 10%

March lab savings - SAVE 10%

March lab savings - SAVE 10%

March lab savings - SAVE 10%

March lab savings - SAVE 10%

March lab savings - SAVE 10%

March lab savings - SAVE 10%

March lab savings - SAVE 10%

March lab savings - SAVE 10%

March lab savings - SAVE 10%

March lab savings - SAVE 10%

March lab savings - SAVE 10%

March lab savings - SAVE 10%

March lab savings - SAVE 10%

March lab savings - SAVE 10%

Preclinical services

Preclinical services

Preclinical services

Preclinical services

Preclinical services

Preclinical services

Preclinical services

Preclinical services

Preclinical services

T-SPOT.TB testing services.

T-SPOT.TB testing services.

T-SPOT.TB testing services.

T-SPOT.TB testing services.

T-SPOT.TB testing services.

T-SPOT.TB testing services.

T-SPOT.TB testing services.

T-SPOT.TB testing services.

T-SPOT.TB testing services.

T-SPOT.TB testing services.

T-SPOT.TB testing services.

T-SPOT.TB testing services.

T-SPOT.TB testing services.

CHOSOURCE expression platform

CHOSOURCE expression platform

CHOSOURCE expression platform

CHOSOURCE expression platform

CHOSOURCE expression platform

T-SPOT.TB testing services.

T-SPOT.TB testing services.

T-SPOT.TB testing services.

T-SPOT.TB testing services.

T-SPOT.TB testing services.

T-SPOT.TB testing services.

T-SPOT.TB testing services.

T-SPOT.TB testing services.

T-SPOT.TB testing services.

T-SPOT.TB testing services.

T-SPOT.TB testing services.

T-SPOT.TB testing services.

March lab savings - SAVE 10%

March lab savings - SAVE 10%

March lab savings - SAVE 10%