Novogene’s Whole Transcriptome Sequencing service equips the researchers with cutting-edge NGS solutions that provide in-depth bioinformatic analyses of all RNA transcripts including mRNA and non-coding RNA (lncRNA, sRNA, and circRNA). By using this paired-end sequencing approach, we can precisely quantify gene and transcript levels, identify splice variants and novel characteristics of the transcriptome. This competitive approach investigates and explores potential transcriptional and regulatory network mechanisms while providing key insights into the interaction functionalities from a comprehensive transcriptomic perspective.
Applications
Whole Transcriptome Sequencing finds its applications in:
- Profiling mRNA and ncRNA in a single run
- Exploring miRNA sponge and target regulatory elements
- Investigating regulatory networks among lncRNA/circRNA-miRNA-gene pairs
Benefits
- Whole Transcriptome Sequencing provides a more comprehensive analysis of transcriptional regulation network, compared to mRNA-seq, lncRNA-seq, sRNA-seq, and circRNA-seq, respectively.
- Whole transcriptome sequencing helps researchers in identifying biomarkers across a wide transcript range.
- WTS allows the capture of both known and new features.
- It enables the whole transcriptome profiling across a broad dynamic scale.
Specifications: RNA Sample Requirements
| Library Type | Sample Type | Amount | RNA Integrity Number (Agilent 2100) |
Purity (NanoDrop) |
| lncRNA Library & small RNA Library | Total RNA | ≥ 2.5 μg | Animal ≥ 7.5, Plant ≥ 7, with smooth baseline | A260/280 = 1.8-2.2; A260/230 ≥ 1.8; |
| lncRNA library & small RNA library & circRNA library | ≥ 4.5 μg |
Specifications: Sequencing and Analysis
| Sequencing Platform | Illumina NovaSeq 6000 Sequencing System |
| Read length | Paired-end 150bp for lncRNA/circRNA library
Single-end 50bp for small RNA library |
| Recommended Data Amount | ≥ 40 million read pairs per sample (lncRNA library); 10-20 million reads per sample (small RNA library); |
| Content of Data Analysis | Individual ncRNA analysis package
Network analysis between lncRNA vs miRNA, mRNA vs miRNA, circRNA vs miRNA, lncRNA-miRNA-mRNA, and circRNA-miRNA-mRNA |
Project Workflow
The first step of the project workflow includes the 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 subsequently tested for its quality (Library QC). Next, a paired-end 150 bp sequencing strategy is used to sequence the lncRNA and circRNA library and single-end 50bp is used to sequence the small RNA library. The resulting data go through quality data control (Data QC) to guarantee the quality of the resulting data. Finally, bioinformatic analyses are performed and publication-ready results are provided. The following flowsheet describes the step-by-step protocol.
