This method is powered by Nanopore technology. It captures the entire transcripts from the 5’ UTR to the 3’ poly-A tail directly, allowing researchers to capture the true structure, poly-A tail lengths, and modifications of transcripts such as 5-methylcytosine (5mC) and N6-methyladenosine (m6A). The ability to detect post-transcriptional modifications opens new avenues in understanding RNA biology, particularly in studying regulatory elements and the functional dynamics of the transcriptome.
Applications
- Investigate the role of RNA modifications on gene regulation and disease mechanisms.
- Estimate poly-A tail length to study post-transcriptional gene expression regulation, such as translation efficiency, RNA stability, cellular responses, and disease-related dysregulation.
- Identify novel isoforms to understand alternative splicing, polyadenylation, and RNA editing, revealing gene expression complexity.
- Profiling transcript abundance across different tissues, conditions, or treatments to support studies on gene expression regulation.
Benefits
- Native RNA sequencing: Avoids artifacts introduced during reverse transcription or amplification, offering more accurate sequencing results.
- Direct detection of RNA modifications: Captures important RNA modifications like methylation that can play a crucial role in gene regulation.
- Long-read sequencing: Provides full-length RNA reads, enabling the discovery of novel isoforms and detailed analysis of transcript variants.
- Comprehensive transcriptome profiling: Offers deeper insights into complex transcriptomes, supporting research in disease mechanisms, gene expression, and RNA biology.
Specifications: RNA Sample Requirements
| Library Type | Sample Type | Amount* | RNA Integrity Number (Agilent 2100) |
Purity (NanoDrop) |
| Nanopore Direct RNA library | Total RNA (Animal) |
≥ 20 μg | ≥ 6.5, with smooth base line | A260/280 = 1.8-2.2 A260/230 ≥ 1.3-2.5 |
| Total RNA (Plant & Fungus) |
≥ 50 μg | ≥ 6.5, with smooth base line | A260/280 = 1.8-2.2 A260/230 ≥ 1.3-2.5 |
*The starting material is total RNA prior to poly(A) enrichment.
Contact us for more information regarding samples that do not meet the minimum sample requirements.
Specifications: Sequencing and Analysis
| Sequencing Platform | Nanopore PromethION |
| Sequencing depth | 10M~20M reads/sample (1 PromethION cell/ sample) |
| Customized Data Analysis | Full-length transcripts identification and classification
Transcript structure analysis Alternative splicing Novel gene/ Novel transcript prediction and annotation Transcription factor analysis LncRNA prediction Quantification analysis Quantification analysis Differential gene expression analysis GO/KEGG enrichment analysis of differentially expressed genes RNA modification analysis Detection of methylation sites and motif analysis (5mC/ m6A) KEGG/GO enrichment analysis Differential methylation site analysis Poly(A) analysis |
Project Workflow
Novogene provides high-quality products and expert services throughout the entire project workflow. Every step is carefully designed and executed to meet rigorous scientific standards, ensuring exceptional research outcomes. To guarantee the accuracy and reliability of sequencing data, stringent quality control (QC) measures are implemented at each stage of the process. The workflow encompasses key steps such as sample preparation and quantification, fragmentation and library preparation, library quality control, sequencing, and bioinformatics analysis.
