Small RNA Sequencing (sRNA-seq)

Profiling miRNAs and other short regulatory RNAs to explore gene regulation, biomarker discovery and biological response.

Small RNA sequencing, also known as small RNA-seq or sRNA-seq, enables the detection and quantification of short regulatory RNA molecules involved in post-transcriptional gene regulation. It is commonly used to profile miRNAs and other small RNA species, supporting studies of gene regulation, biomarker discovery and biological response.

Novogene Europe supports small RNA-seq projects using qualified RNA inputs and library preparation workflows designed to enrich short RNA molecules for sequencing. Bioinformatics analysis supports small RNA identification, classification, quantification and differential expression analysis, helping researchers compare small RNA profiles across sample groups or experimental conditions.

Service Highlights

miRNA and Small RNA Profiling

Profile miRNAs and other short regulatory RNA molecules involved in post-transcriptional regulation, biological response and gene expression control.

Size-selected Small RNA Libraries

Library preparation workflows enrich short RNA molecules for sequencing, supporting focused analysis of miRNAs and other small regulatory RNA species.

Small RNA Identification and Prediction

Bioinformatics workflows support classification and quantification of known small RNAs, with prediction of potential novel small RNA candidates.

Differential Expression and Target Insight

Compare small RNA expression across sample groups and explore predicted targets, pathways and biological functions linked to expression changes.

Applications

Gene Regulation and Regulatory Networks

Small RNA-seq helps researchers study miRNA-mediated gene regulation and explore how small RNA expression patterns influence biological pathways and cellular processes.

Disease and Biomarker Research

Small RNA profiles can support research into disease mechanisms, treatment response and biomarker discovery across tissue, biofluid or exosomal RNA samples.

Development and Cell State Studies

Small RNA-seq supports studies of development, differentiation and cell-state changes by comparing regulatory RNA expression across stages, tissues or conditions.

Stress and Environmental Response

In plant, animal and microbial-associated studies, small RNA profiling can help investigate regulatory responses to stress, infection, environmental change or experimental treatment.

Specifications

Sample Requirements

The quantities listed are provided for reference only. Please download the Sample Requirements for detailed guidance, or contact us to discuss your project-specific requirements.

Library Type	Sample Type	Amount	RNA Integrity Number (Agilent 5400)	Purity (NanoDrop)
Small RNA Library	Total RNA	≥ 1 μg	Animal ≥ 7.5, Plant ≥ 7, with smooth baseline	OD260/280≥ 2.0; OD260/230≥ 2.0; no degradation, no contamination
Exosomal Small RNA Library	Exosomal RNA	≥ 10 ng	Fragments between 25- 40nt, FU*>10, with flat baseline	OD260/280≥2.0. OD260/230≥2.0. no degradation, no contamination

Sample Requirements

The quantities listed are provided for reference only. Please download the Sample Requirements for detailed guidance, or contact us to discuss your project-specific requirements.

Library Type	Sample Type	Amount	RNA Integrity Number (Agilent 5400)	Purity (NanoDrop)
Small RNA Library	Total RNA	≥ 1 μg	Animal ≥ 7.5, Plant ≥ 7, with smooth baseline	OD260/280≥ 2.0; OD260/230≥ 2.0; no degradation, no contamination
Exosomal Small RNA Library	Exosomal RNA	≥ 10 ng	Fragments between 25- 40nt, FU*>10, with flat baseline	OD260/280≥2.0. OD260/230≥2.0. no degradation, no contamination

Sample Requirements

The quantities listed are provided for reference only. Please download the Sample Requirements for detailed guidance, or contact us to discuss your project-specific requirements.

Library Type	Sample Type	Amount	RNA Integrity Number (Agilent 5400)	Purity (NanoDrop)
Small RNA Library	Total RNA	≥ 1 μg	Animal ≥ 7.5, Plant ≥ 7, with smooth baseline	OD260/280≥ 2.0; OD260/230≥ 2.0; no degradation, no contamination
Exosomal Small RNA Library	Exosomal RNA	≥ 10 ng	Fragments between 25- 40nt, FU*>10, with flat baseline	OD260/280≥2.0. OD260/230≥2.0. no degradation, no contamination

Sample Requirements

The quantities listed are provided for reference only. Please download the Sample Requirements for detailed guidance, or contact us to discuss your project-specific requirements.

Library Type	Sample Type	Amount	RNA Integrity Number (Agilent 5400)	Purity (NanoDrop)
Small RNA Library	Total RNA	≥ 1 μg	Animal ≥ 7.5, Plant ≥ 7, with smooth baseline	OD260/280≥ 2.0; OD260/230≥ 2.0; no degradation, no contamination
Exosomal Small RNA Library	Exosomal RNA	≥ 10 ng	Fragments between 25- 40nt, FU*>10, with flat baseline	OD260/280≥2.0. OD260/230≥2.0. no degradation, no contamination

Sequencing and Analysis

Recommended data outputs and analysis examples are provided for reference only. For detailed information, please contact us to discuss your project-specific analysis requirements.

Sequencing Platform	Illumina NovaSeq X Plus
Read Length	Single-end 50 bp format data
Recommended Sequencing Depth	≥ 10-20 M read pairs per sample
Standard Analysis (miRNA)	Data Quality Control Summary of Length Distribution Common & Specific Sequence Summary Identification & Characterization of miRNAs Classification & Annotation of miRNA Quantification & Differential Expression Analysis Functional Enrichment Analysis

Sequencing and Analysis

Recommended data outputs and analysis examples are provided for reference only. For detailed information, please contact us to discuss your project-specific analysis requirements.

Sequencing Platform	Illumina NovaSeq X Plus
Read Length	Single-end 50 bp format data
Recommended Sequencing Depth	≥ 10-20 M read pairs per sample
Standard Analysis (miRNA)	Data Quality Control Summary of Length Distribution Common & Specific Sequence Summary Identification & Characterization of miRNAs Classification & Annotation of miRNA Quantification & Differential Expression Analysis Functional Enrichment Analysis

Sequencing and Analysis

Recommended data outputs and analysis examples are provided for reference only. For detailed information, please contact us to discuss your project-specific analysis requirements.

Sequencing Platform	Illumina NovaSeq X Plus
Read Length	Single-end 50 bp format data
Recommended Sequencing Depth	≥ 10-20 M read pairs per sample
Standard Analysis (miRNA)	Data Quality Control Summary of Length Distribution Common & Specific Sequence Summary Identification & Characterization of miRNAs Classification & Annotation of miRNA Quantification & Differential Expression Analysis Functional Enrichment Analysis

Sequencing and Analysis

Recommended data outputs and analysis examples are provided for reference only. For detailed information, please contact us to discuss your project-specific analysis requirements.

Sequencing Platform	Illumina NovaSeq X Plus
Read Length	Single-end 50 bp format data
Recommended Sequencing Depth	≥ 10-20 M read pairs per sample
Standard Analysis (miRNA)	Data Quality Control Summary of Length Distribution Common & Specific Sequence Summary Identification & Characterization of miRNAs Classification & Annotation of miRNA Quantification & Differential Expression Analysis Functional Enrichment Analysis

Specifications

Sample Requirements

The quantities listed are provided for reference only. Please download the Sample Requirements for detailed guidance, or contact us to discuss your project-specific requirements.

Library Type	Sample Type	Amount	RNA Integrity Number (Agilent 5400)	Purity (NanoDrop)
Small RNA Library	Total RNA	≥ 1 μg	Animal ≥ 7.5, Plant ≥ 7, with smooth baseline	OD260/280≥ 2.0; OD260/230≥ 2.0; no degradation, no contamination
Exosomal Small RNA Library	Exosomal RNA	≥ 10 ng	Fragments between 25- 40nt, FU*>10, with flat baseline	OD260/280≥2.0. OD260/230≥2.0. no degradation, no contamination

Sample Requirements

The quantities listed are provided for reference only. Please download the Sample Requirements for detailed guidance, or contact us to discuss your project-specific requirements.

Library Type	Sample Type	Amount	RNA Integrity Number (Agilent 5400)	Purity (NanoDrop)
Small RNA Library	Total RNA	≥ 1 μg	Animal ≥ 7.5, Plant ≥ 7, with smooth baseline	OD260/280≥ 2.0; OD260/230≥ 2.0; no degradation, no contamination
Exosomal Small RNA Library	Exosomal RNA	≥ 10 ng	Fragments between 25- 40nt, FU*>10, with flat baseline	OD260/280≥2.0. OD260/230≥2.0. no degradation, no contamination

Sample Requirements

The quantities listed are provided for reference only. Please download the Sample Requirements for detailed guidance, or contact us to discuss your project-specific requirements.

Library Type	Sample Type	Amount	RNA Integrity Number (Agilent 5400)	Purity (NanoDrop)
Small RNA Library	Total RNA	≥ 1 μg	Animal ≥ 7.5, Plant ≥ 7, with smooth baseline	OD260/280≥ 2.0; OD260/230≥ 2.0; no degradation, no contamination
Exosomal Small RNA Library	Exosomal RNA	≥ 10 ng	Fragments between 25- 40nt, FU*>10, with flat baseline	OD260/280≥2.0. OD260/230≥2.0. no degradation, no contamination

Sample Requirements

The quantities listed are provided for reference only. Please download the Sample Requirements for detailed guidance, or contact us to discuss your project-specific requirements.

Library Type	Sample Type	Amount	RNA Integrity Number (Agilent 5400)	Purity (NanoDrop)
Small RNA Library	Total RNA	≥ 1 μg	Animal ≥ 7.5, Plant ≥ 7, with smooth baseline	OD260/280≥ 2.0; OD260/230≥ 2.0; no degradation, no contamination
Exosomal Small RNA Library	Exosomal RNA	≥ 10 ng	Fragments between 25- 40nt, FU*>10, with flat baseline	OD260/280≥2.0. OD260/230≥2.0. no degradation, no contamination

Sequencing and Analysis

Recommended data outputs and analysis examples are provided for reference only. For detailed information, please contact us to discuss your project-specific analysis requirements.

Sequencing Platform	Illumina NovaSeq X Plus
Read Length	Single-end 50 bp format data
Recommended Sequencing Depth	≥ 10-20 M read pairs per sample
Standard Analysis (miRNA)	Data Quality Control Summary of Length Distribution Common & Specific Sequence Summary Identification & Characterization of miRNAs Classification & Annotation of miRNA Quantification & Differential Expression Analysis Functional Enrichment Analysis

Sequencing and Analysis

Recommended data outputs and analysis examples are provided for reference only. For detailed information, please contact us to discuss your project-specific analysis requirements.

Sequencing Platform	Illumina NovaSeq X Plus
Read Length	Single-end 50 bp format data
Recommended Sequencing Depth	≥ 10-20 M read pairs per sample
Standard Analysis (miRNA)	Data Quality Control Summary of Length Distribution Common & Specific Sequence Summary Identification & Characterization of miRNAs Classification & Annotation of miRNA Quantification & Differential Expression Analysis Functional Enrichment Analysis

Sequencing and Analysis

Recommended data outputs and analysis examples are provided for reference only. For detailed information, please contact us to discuss your project-specific analysis requirements.

Sequencing Platform	Illumina NovaSeq X Plus
Read Length	Single-end 50 bp format data
Recommended Sequencing Depth	≥ 10-20 M read pairs per sample
Standard Analysis (miRNA)	Data Quality Control Summary of Length Distribution Common & Specific Sequence Summary Identification & Characterization of miRNAs Classification & Annotation of miRNA Quantification & Differential Expression Analysis Functional Enrichment Analysis

Sequencing and Analysis

Recommended data outputs and analysis examples are provided for reference only. For detailed information, please contact us to discuss your project-specific analysis requirements.

Sequencing Platform	Illumina NovaSeq X Plus
Read Length	Single-end 50 bp format data
Recommended Sequencing Depth	≥ 10-20 M read pairs per sample
Standard Analysis (miRNA)	Data Quality Control Summary of Length Distribution Common & Specific Sequence Summary Identification & Characterization of miRNAs Classification & Annotation of miRNA Quantification & Differential Expression Analysis Functional Enrichment Analysis

Resources

Example Analysis Output

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Small RNA Length Distribution

Review sequence length distribution to assess small RNA size profiles and enrichment patterns.

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Repeat Classification

Summarise unique reads aligned to repeat classes and other annotated RNA categories.

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miRNA TPM Distribution

Compare TPM expression distributions across samples to review overall miRNA abundance profiles.

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Differential miRNA Heatmap

Visualise expression patterns of differentially expressed miRNAs across samples and experimental groups.

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Target Gene Enrichment

Identify enriched functional terms associated with predicted miRNA target genes for downstream interpretation.

Webinars

ncRNA-seq Results Explained: What you can explore with non-coding RNA data

Learn how to navigate and interpret non-coding RNA sequencing (ncRNA-seq) data, including lncRNA-seq, small RNA-seq, and circRNA-seq. This webinar will provide an overview of analysis workflows, data visualisation approaches, and the tools needed to transform complex datasets into meaningful biological insights.

Learn More

Frequently Asked Questions

What is small RNA sequencing?

Small RNA-seq is a sequencing approach for profiling short non-coding RNAs such as miRNAs and other small RNA species.

Which small RNAs can sRNA-seq detect and quantify?

How do miRNAs differ from other sRNAs?

What research applications use small RNA-seq?

Can small RNA-seq identify novel small RNAs?

Explore Transcriptomics Insights

Access webinars, brochures and case studies covering sequencing technologies, study planning and data interpretation.

Speak with a Specialist

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