At Novogene, cDNA libraries are sequenced using the state-of-the-art Illumina NovaSeq platforms, which utilize a paired-end 150 bp sequencing strategy (short-reads). Along with our rich experience and strong sequencing capacity, Novogene offers services to meet a wide array of research objectives. Our services not only include eukaryotic mRNA sequencing (mRNA-seq) services, Novogene can also deliver data on prokaryotic transcripts, non-coding RNAs, full-length isoforms (long-reads), whole transcriptomes, and meta-transcriptomes.
Applications
- mRNA-seq is a powerful tool to analyze the cell transcriptome profile. Novogene’s professional services help on research goals in a wide range of applications, including:
- Quantitative profiling of transcripts in different tissues or samples, under various conditions and treatments
- Discovery of novel transcripts, alternative splicing (AS), and transcript variations
- Research of developmental mechanisms and drug resistance through tissue-specific transcripts or time-course gene expression
- Biomarker discovery based on novel transcripts/isoforms, SNP/InDel identification, and fusion gene analysis
- Omics analysis in combination with the transcriptome
- Investigation of pathogenic mechanisms and clinical subtypes in clinical diagnosis
Benefits
Novogene mRNA-seq offers high throughput and high accuracy (with Q30 score ≥ 85%) coupled with a low initial RNA input required. Novogene has extensive experience providing RNA-Seq services, having successfully completed thousands of projects to help multiple researchers to publish in high impact factor journals.
Novogene offers inclusive solutions for quantification, differential gene expression, annotation of novel transcripts, alternative splicing, discovery of fusion gene, and other potential variations. Highly-qualified bioinformaticians deliver publication-ready data using personalized pipelines for species either with or without a reference genome.
Specifications: Sample Requirements
| Library Type | Sample Type | Amount | RNA Integrity Number (Agilent 2100) | Purity (NanoDrop) |
| Eukaryotic RNA-Seq (cDNA library) | Total RNA | ≥ 200 ng | ≥ 4.0, with smooth base line | A260/280 = 1.8-2.2 A260/230 ≥ 1.8 |
| Total RNA (Blood) | ≥ 400 ng | ≥ 5.8, with smooth base line | ||
| Amplified cDNA (double-stranded) | ≥ 100 ng | Fragments between 400bp and 5000bp with main peak at ~2000bp | A260/280 = 1.8-2.0 A260/230 ≥ 1.8 |
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| Eukaryotic RNA-Seq (strand specific library) | Total RNA | ≥ 400 ng | ≥ 5.8, with smooth base line | A260/280 = 1.8-2.2 A260/230 ≥ 1.8 |
Specifications: Sequencing and Analysis
| Sequencing Platform | Illumina NovaSeq 6000 Sequencing System |
| 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
The project workflow starts with sample quality control (Sample QC) to ensure that your samples meet the criteria of the RNA-Seq technique. Then, the appropriate library is prepared according to your target organism and application, and subsequently tested for its quality (Library QC). Next, a 150 bp paired-end sequencing strategy is used to sequence the samples and the quality of the resulting data is also checked for its quality (Data QC). Finally, bioinformatic analyses are performed and publication-ready results are provided. The following flowsheet describes the step-by-step protocol our mRNA-seq technique follows.
Preparation of sample is followed by the RNA library preparation. RNA library is formed by polyA capture (or rRNA removal) and reverse transcription of cDNA. Illumina PE150 technology is employed to sequence the sample and the final stage involves the bioinformatics analysis.
