mRNA Sequencing

Genome-wide profiling of messenger RNA expression to reveal coding transcript dynamics and gene regulatory changes across biological conditions.

Messenger RNA sequencing (mRNA-seq) has revolutionized the exploration of cellular functionality, offering researchers unparalleled insights into the transcriptional landscape of cells. By leveraging high-throughput and precise next-generation sequencing (NGS) techniques, RNA-seq unveils gene expression profiles and highlights the dynamic variations within the transcriptome. This innovative technique selectively captures or enriches single-stranded messenger RNAs (mRNAs), converting them into complementary DNA (cDNA) for streamlined library preparation.

At Novogene, we employ cutting-edge Illumina NovaSeq platforms for sequencing cDNA libraries. These platforms utilize a paired-end 150 bp sequencing strategy, providing high-quality short-read data. Leveraging our extensive experience and robust sequencing capacity, Novogene offers a diverse range of services to meet various research objectives. Our offerings extend beyond eukaryotic mRNA sequencing (mRNA-seq). Novogene can also deliver data on prokaryotic transcripts, non-coding RNAs, full-length isoforms (long-reads), whole transcriptomes, and meta-transcriptomes.

Why Choose Novogene for Your mRNA-seq Needs?

Precision and Efficiency

Consistently exceeding manufacturer performance benchmarks.

Local Support

Regional sequencing facilities and dedicated customer service.

Fast Turnaround

From QC to FASTQ Files in As Little As 7 Business Days.

Proven Expertise

Over 1 million RNA-seq samples delivered annually.

Comprehensive Solutions

Customized workflows and analysis for challenging or complex samples.

Expert Bioinformatics

Expert bioinformaticians and well-established bioinformatics pipelines ensure publication-ready data of the highest quality.

Unlocking the Potential of mRNA Sequencing

Discover the power of mRNA-seq with Novogene’s services, designed to assist in a variety of research goals:

Explore Diversity

Understand transcript profiles across different tissues, conditions, and treatments.

Uncover Novel Insights

Identify new transcripts, alternative splicing, and variations for a comprehensive view.

Navigate Development

Study developmental mechanisms and drug resistance through time-course gene expression.

Find Biomarkers

Discover potential biomarkers using novel transcripts, SNP/InDel identification, and fusion gene analysis.

Comprehensive Analysis

Combine transcriptome data with omics analysis for a holistic perspective.

Clinical Insight

Investigate pathogenic mechanisms and clinical subtypes for precise clinical diagnosis.

Specifications

Sample Requirements

Sample amounts are listed for reference only. Download the Sample Submission Guidelines to learn more. For detailed information, please contact us with your customized requests.

Service	Sample Type	Amount (Qubit®)	Volume	Concentration	RIN (Agilent 5400)	Purity (NanoDrop™)
Eukaryotic mRNA (polyA enrichment)	Total RNA (animal, plant, fungus)	≥100 ng	≥10 μL	≥10 ng/μL	≥4.0, with flat baseline	OD260/280 = 1.8-2.2; OD260/230 ≥ 2.0; no degradation, no contamination
Eukaryotic mRNA (polyA enrichment)	Total RNA (blood)	≥400 ng	≥20 μL	≥20 ng/μL	≥5.0, with flat baseline
Strand-specific Eukaryotic mRNA (polyA enrichment)	Total RNA (animal, plant, fungus)	≥200 ng	≥20 μL	≥20 ng/μL	≥5.0, with flat baseline
Strand-specific Eukaryotic mRNA (polyA enrichment)	Total RNA (blood)	≥400 ng	≥20 μL	≥20 ng/μL	≥5.0, with flat baseline

Sample Requirements

Sample amounts are listed for reference only. Download the Sample Submission Guidelines to learn more. For detailed information, please contact us with your customized requests.

Service	Sample Type	Amount (Qubit®)	Volume	Concentration	RIN (Agilent 5400)	Purity (NanoDrop™)
Eukaryotic mRNA (polyA enrichment)	Total RNA (animal, plant, fungus)	≥100 ng	≥10 μL	≥10 ng/μL	≥4.0, with flat baseline	OD260/280 = 1.8-2.2; OD260/230 ≥ 2.0; no degradation, no contamination
Eukaryotic mRNA (polyA enrichment)	Total RNA (blood)	≥400 ng	≥20 μL	≥20 ng/μL	≥5.0, with flat baseline
Strand-specific Eukaryotic mRNA (polyA enrichment)	Total RNA (animal, plant, fungus)	≥200 ng	≥20 μL	≥20 ng/μL	≥5.0, with flat baseline
Strand-specific Eukaryotic mRNA (polyA enrichment)	Total RNA (blood)	≥400 ng	≥20 μL	≥20 ng/μL	≥5.0, with flat baseline

Sample Requirements

Sample amounts are listed for reference only. Download the Sample Submission Guidelines to learn more. For detailed information, please contact us with your customized requests.

Service	Sample Type	Amount (Qubit®)	Volume	Concentration	RIN (Agilent 5400)	Purity (NanoDrop™)
Eukaryotic mRNA (polyA enrichment)	Total RNA (animal, plant, fungus)	≥100 ng	≥10 μL	≥10 ng/μL	≥4.0, with flat baseline	OD260/280 = 1.8-2.2; OD260/230 ≥ 2.0; no degradation, no contamination
Eukaryotic mRNA (polyA enrichment)	Total RNA (blood)	≥400 ng	≥20 μL	≥20 ng/μL	≥5.0, with flat baseline
Strand-specific Eukaryotic mRNA (polyA enrichment)	Total RNA (animal, plant, fungus)	≥200 ng	≥20 μL	≥20 ng/μL	≥5.0, with flat baseline
Strand-specific Eukaryotic mRNA (polyA enrichment)	Total RNA (blood)	≥400 ng	≥20 μL	≥20 ng/μL	≥5.0, with flat baseline

Sample Requirements

Sample amounts are listed for reference only. Download the Sample Submission Guidelines to learn more. For detailed information, please contact us with your customized requests.

Service	Sample Type	Amount (Qubit®)	Volume	Concentration	RIN (Agilent 5400)	Purity (NanoDrop™)
Eukaryotic mRNA (polyA enrichment)	Total RNA (animal, plant, fungus)	≥100 ng	≥10 μL	≥10 ng/μL	≥4.0, with flat baseline	OD260/280 = 1.8-2.2; OD260/230 ≥ 2.0; no degradation, no contamination
Eukaryotic mRNA (polyA enrichment)	Total RNA (blood)	≥400 ng	≥20 μL	≥20 ng/μL	≥5.0, with flat baseline
Strand-specific Eukaryotic mRNA (polyA enrichment)	Total RNA (animal, plant, fungus)	≥200 ng	≥20 μL	≥20 ng/μL	≥5.0, with flat baseline
Strand-specific Eukaryotic mRNA (polyA enrichment)	Total RNA (blood)	≥400 ng	≥20 μL	≥20 ng/μL	≥5.0, with flat baseline

Sequencing and Analysis

Recommended data outputs and analysis contents displayed are for reference only. For detailed information, please contact us with your customized requests.

Sequencing Platform	Illumina NovaSeq X-Plus Sequencing Platform
Read Length	Paired-end 150 bp
Data Output	≥ 20 million read pairs per sample for species with reference genome ≥ 50 million read pairs per sample for species without reference genome (de novo transcriptome assembly projects)
Data Analysis Capability	Data Quality Control Gene expression quantification Differential expression profiling Functional enrichment analysis Novel transcripts identification SNP & InDel analysis Alternative splicing (AS) analysis Fusion gene prediction Protein-Protein Interaction (PPI) analysis Transcription factors and oncogene functional annotation

Project Workflow

Our mRNA-Seq workflow ensures high-quality results through a rigorous, step-by-step process:

Sequencing and Analysis

Recommended data outputs and analysis contents displayed are for reference only. For detailed information, please contact us with your customized requests.

Sequencing Platform	Illumina NovaSeq X-Plus Sequencing Platform
Read Length	Paired-end 150 bp
Data Output	≥ 20 million read pairs per sample for species with reference genome ≥ 50 million read pairs per sample for species without reference genome (de novo transcriptome assembly projects)
Data Analysis Capability	Data Quality Control Gene expression quantification Differential expression profiling Functional enrichment analysis Novel transcripts identification SNP & InDel analysis Alternative splicing (AS) analysis Fusion gene prediction Protein-Protein Interaction (PPI) analysis Transcription factors and oncogene functional annotation

Project Workflow

Our mRNA-Seq workflow ensures high-quality results through a rigorous, step-by-step process:

Sequencing and Analysis

Recommended data outputs and analysis contents displayed are for reference only. For detailed information, please contact us with your customized requests.

Sequencing Platform	Illumina NovaSeq X-Plus Sequencing Platform
Read Length	Paired-end 150 bp
Data Output	≥ 20 million read pairs per sample for species with reference genome ≥ 50 million read pairs per sample for species without reference genome (de novo transcriptome assembly projects)
Data Analysis Capability	Data Quality Control Gene expression quantification Differential expression profiling Functional enrichment analysis Novel transcripts identification SNP & InDel analysis Alternative splicing (AS) analysis Fusion gene prediction Protein-Protein Interaction (PPI) analysis Transcription factors and oncogene functional annotation

Project Workflow

Our mRNA-Seq workflow ensures high-quality results through a rigorous, step-by-step process:

Sequencing and Analysis

Recommended data outputs and analysis contents displayed are for reference only. For detailed information, please contact us with your customized requests.

Sequencing Platform	Illumina NovaSeq X-Plus Sequencing Platform
Read Length	Paired-end 150 bp
Data Output	≥ 20 million read pairs per sample for species with reference genome ≥ 50 million read pairs per sample for species without reference genome (de novo transcriptome assembly projects)
Data Analysis Capability	Data Quality Control Gene expression quantification Differential expression profiling Functional enrichment analysis Novel transcripts identification SNP & InDel analysis Alternative splicing (AS) analysis Fusion gene prediction Protein-Protein Interaction (PPI) analysis Transcription factors and oncogene functional annotation

Project Workflow

Our mRNA-Seq workflow ensures high-quality results through a rigorous, step-by-step process:

Specifications

Sample Requirements

Sample amounts are listed for reference only. Download the Sample Submission Guidelines to learn more. For detailed information, please contact us with your customized requests.

Service	Sample Type	Amount (Qubit®)	Volume	Concentration	RIN (Agilent 5400)	Purity (NanoDrop™)
Eukaryotic mRNA (polyA enrichment)	Total RNA (animal, plant, fungus)	≥100 ng	≥10 μL	≥10 ng/μL	≥4.0, with flat baseline	OD260/280 = 1.8-2.2; OD260/230 ≥ 2.0; no degradation, no contamination
Eukaryotic mRNA (polyA enrichment)	Total RNA (blood)	≥400 ng	≥20 μL	≥20 ng/μL	≥5.0, with flat baseline
Strand-specific Eukaryotic mRNA (polyA enrichment)	Total RNA (animal, plant, fungus)	≥200 ng	≥20 μL	≥20 ng/μL	≥5.0, with flat baseline
Strand-specific Eukaryotic mRNA (polyA enrichment)	Total RNA (blood)	≥400 ng	≥20 μL	≥20 ng/μL	≥5.0, with flat baseline

Sample Requirements

Sample amounts are listed for reference only. Download the Sample Submission Guidelines to learn more. For detailed information, please contact us with your customized requests.

Service	Sample Type	Amount (Qubit®)	Volume	Concentration	RIN (Agilent 5400)	Purity (NanoDrop™)
Eukaryotic mRNA (polyA enrichment)	Total RNA (animal, plant, fungus)	≥100 ng	≥10 μL	≥10 ng/μL	≥4.0, with flat baseline	OD260/280 = 1.8-2.2; OD260/230 ≥ 2.0; no degradation, no contamination
Eukaryotic mRNA (polyA enrichment)	Total RNA (blood)	≥400 ng	≥20 μL	≥20 ng/μL	≥5.0, with flat baseline
Strand-specific Eukaryotic mRNA (polyA enrichment)	Total RNA (animal, plant, fungus)	≥200 ng	≥20 μL	≥20 ng/μL	≥5.0, with flat baseline
Strand-specific Eukaryotic mRNA (polyA enrichment)	Total RNA (blood)	≥400 ng	≥20 μL	≥20 ng/μL	≥5.0, with flat baseline

Sample Requirements

Sample amounts are listed for reference only. Download the Sample Submission Guidelines to learn more. For detailed information, please contact us with your customized requests.

Service	Sample Type	Amount (Qubit®)	Volume	Concentration	RIN (Agilent 5400)	Purity (NanoDrop™)
Eukaryotic mRNA (polyA enrichment)	Total RNA (animal, plant, fungus)	≥100 ng	≥10 μL	≥10 ng/μL	≥4.0, with flat baseline	OD260/280 = 1.8-2.2; OD260/230 ≥ 2.0; no degradation, no contamination
Eukaryotic mRNA (polyA enrichment)	Total RNA (blood)	≥400 ng	≥20 μL	≥20 ng/μL	≥5.0, with flat baseline
Strand-specific Eukaryotic mRNA (polyA enrichment)	Total RNA (animal, plant, fungus)	≥200 ng	≥20 μL	≥20 ng/μL	≥5.0, with flat baseline
Strand-specific Eukaryotic mRNA (polyA enrichment)	Total RNA (blood)	≥400 ng	≥20 μL	≥20 ng/μL	≥5.0, with flat baseline

Sample Requirements

Sample amounts are listed for reference only. Download the Sample Submission Guidelines to learn more. For detailed information, please contact us with your customized requests.

Service	Sample Type	Amount (Qubit®)	Volume	Concentration	RIN (Agilent 5400)	Purity (NanoDrop™)
Eukaryotic mRNA (polyA enrichment)	Total RNA (animal, plant, fungus)	≥100 ng	≥10 μL	≥10 ng/μL	≥4.0, with flat baseline	OD260/280 = 1.8-2.2; OD260/230 ≥ 2.0; no degradation, no contamination
Eukaryotic mRNA (polyA enrichment)	Total RNA (blood)	≥400 ng	≥20 μL	≥20 ng/μL	≥5.0, with flat baseline
Strand-specific Eukaryotic mRNA (polyA enrichment)	Total RNA (animal, plant, fungus)	≥200 ng	≥20 μL	≥20 ng/μL	≥5.0, with flat baseline
Strand-specific Eukaryotic mRNA (polyA enrichment)	Total RNA (blood)	≥400 ng	≥20 μL	≥20 ng/μL	≥5.0, with flat baseline

Sequencing and Analysis

Recommended data outputs and analysis contents displayed are for reference only. For detailed information, please contact us with your customized requests.

Sequencing Platform	Illumina NovaSeq X-Plus Sequencing Platform
Read Length	Paired-end 150 bp
Data Output	≥ 20 million read pairs per sample for species with reference genome ≥ 50 million read pairs per sample for species without reference genome (de novo transcriptome assembly projects)
Data Analysis Capability	Data Quality Control Gene expression quantification Differential expression profiling Functional enrichment analysis Novel transcripts identification SNP & InDel analysis Alternative splicing (AS) analysis Fusion gene prediction Protein-Protein Interaction (PPI) analysis Transcription factors and oncogene functional annotation

Project Workflow

Our mRNA-Seq workflow ensures high-quality results through a rigorous, step-by-step process:

Sequencing and Analysis

Recommended data outputs and analysis contents displayed are for reference only. For detailed information, please contact us with your customized requests.

Sequencing Platform	Illumina NovaSeq X-Plus Sequencing Platform
Read Length	Paired-end 150 bp
Data Output	≥ 20 million read pairs per sample for species with reference genome ≥ 50 million read pairs per sample for species without reference genome (de novo transcriptome assembly projects)
Data Analysis Capability	Data Quality Control Gene expression quantification Differential expression profiling Functional enrichment analysis Novel transcripts identification SNP & InDel analysis Alternative splicing (AS) analysis Fusion gene prediction Protein-Protein Interaction (PPI) analysis Transcription factors and oncogene functional annotation

Project Workflow

Our mRNA-Seq workflow ensures high-quality results through a rigorous, step-by-step process:

Sequencing and Analysis

Recommended data outputs and analysis contents displayed are for reference only. For detailed information, please contact us with your customized requests.

Sequencing Platform	Illumina NovaSeq X-Plus Sequencing Platform
Read Length	Paired-end 150 bp
Data Output	≥ 20 million read pairs per sample for species with reference genome ≥ 50 million read pairs per sample for species without reference genome (de novo transcriptome assembly projects)
Data Analysis Capability	Data Quality Control Gene expression quantification Differential expression profiling Functional enrichment analysis Novel transcripts identification SNP & InDel analysis Alternative splicing (AS) analysis Fusion gene prediction Protein-Protein Interaction (PPI) analysis Transcription factors and oncogene functional annotation

Project Workflow

Our mRNA-Seq workflow ensures high-quality results through a rigorous, step-by-step process:

Sequencing and Analysis

Recommended data outputs and analysis contents displayed are for reference only. For detailed information, please contact us with your customized requests.

Sequencing Platform	Illumina NovaSeq X-Plus Sequencing Platform
Read Length	Paired-end 150 bp
Data Output	≥ 20 million read pairs per sample for species with reference genome ≥ 50 million read pairs per sample for species without reference genome (de novo transcriptome assembly projects)
Data Analysis Capability	Data Quality Control Gene expression quantification Differential expression profiling Functional enrichment analysis Novel transcripts identification SNP & InDel analysis Alternative splicing (AS) analysis Fusion gene prediction Protein-Protein Interaction (PPI) analysis Transcription factors and oncogene functional annotation

Project Workflow

Our mRNA-Seq workflow ensures high-quality results through a rigorous, step-by-step process:

Resources

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Sample Correlation

Correlation heatmap among samples.

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Volcano Plot of Differentially Expressed Genes (DEG)

Differentially up-regulated and differentially down-regulated gene map

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Heatmap of DEG

Cluster diagram of DEGs in samples from different groups

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KEGG Enrichment

KEGG enrichment map of DEG

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Hierarchical Cluster of DEG

The genes within the same cluster exhibit similar trends in expression level changes under different treatment conditions.

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Visualization of Alternative Splicing

The differential AS events identified by the rMATS software

Frequently Asked Questions

How much data do I need for RNA-seq?

The amount of data required for RNA-seq depends on the purpose of the research.

For gene-level differential expression with detection of lowly expressed genes, we recommend sequencing deeper, with at least 30–60 million reads depending on the expression levels (start with 30 million reads is reasonable with a good number of replicates).

For general gene-level differential expression, the ENCODE guidelines suggest 30 million reads per sample (stranded). However, 20 million reads per sample is often sufficient if there are a good number of replicates (>3).

How many repeated samples are required?

When should strand-specific RNA-seq be preferred over non-strand-specific approaches?

What precautions should l take when sending samples for RNA-seq outsourcing?

What is the difference between RNA-seq and single-cell RNA-seq?

More Services

10x Single Cell RNA Sequencing

Full-Length Transcriptome Sequencing

Total RNA Sequencing

Small RNA Sequencing

Metatranscriptome Sequencing

Prokaryotic RNA Sequencing

Ready to start your project?

Our platform offers tailored solutions for your unique experimental needs, ensuring a seamless experience from project design to data delivery.

Service

Sample Type

Amount (Qubit®)

Volume

Concentration

RIN (Agilent 5400)

Purity (NanoDrop™)

Eukaryotic mRNA
(polyA enrichment)

Total RNA (animal, plant, fungus)

≥100 ng

≥10 μL

≥10 ng/μL

≥4.0, with flat baseline

OD260/280 = 1.8-2.2;
OD260/230 ≥ 2.0;
no degradation,
no contamination

Total RNA (blood)

≥400 ng

≥20 μL

≥20 ng/μL

≥5.0, with flat baseline

Strand-specific Eukaryotic mRNA
(polyA enrichment)

Total RNA (animal, plant, fungus)

≥200 ng

≥20 μL

≥20 ng/μL

≥5.0, with flat baseline

Total RNA (blood)

≥400 ng

≥20 μL

≥20 ng/μL

≥5.0, with flat baseline

Sequencing Platform

Illumina NovaSeq X-Plus Sequencing Platform

Read Length

Paired-end 150 bp

Data Output

≥ 20 million read pairs per sample for species with reference genome
≥ 50 million read pairs per sample for species without reference genome (de novo transcriptome assembly projects)

Data Analysis Capability

Data Quality Control
Gene expression quantification
Differential expression profiling
Functional enrichment analysis
Novel transcripts identification
SNP & InDel analysis
Alternative splicing (AS) analysis
Fusion gene prediction
Protein-Protein Interaction (PPI) analysis
Transcription factors and oncogene functional annotation

Service

Sample Type

Amount (Qubit®)

Volume

Concentration

RIN (Agilent 5400)

Purity (NanoDrop™)

Eukaryotic mRNA
(polyA enrichment)

Total RNA (animal, plant, fungus)

≥100 ng

≥10 μL

≥10 ng/μL

≥4.0, with flat baseline

OD260/280 = 1.8-2.2;
OD260/230 ≥ 2.0;
no degradation,
no contamination

Total RNA (blood)

≥400 ng

≥20 μL

≥20 ng/μL

≥5.0, with flat baseline

Strand-specific Eukaryotic mRNA
(polyA enrichment)

Total RNA (animal, plant, fungus)

≥200 ng

≥20 μL

≥20 ng/μL

≥5.0, with flat baseline

Total RNA (blood)

≥400 ng

≥20 μL

≥20 ng/μL

≥5.0, with flat baseline

Sequencing Platform

Illumina NovaSeq X-Plus Sequencing Platform

Read Length

Paired-end 150 bp

Data Output

≥ 20 million read pairs per sample for species with reference genome
≥ 50 million read pairs per sample for species without reference genome (de novo transcriptome assembly projects)

Data Analysis Capability