Overview of Our Nanopore full-length cDNA Sequencing
High-throughput sequencing dramatically changed our view of transcriptome architectures and allowed for ground-breaking discoveries in RNA biology. Transcriptome sequencing (RNA-Seq) is a powerful tool for studying changes in gene expression levels between different experimental groups. However, short-read sequencing technology requires manual fragmentation of transcripts during the library construction process, resulting in many complications in transcriptome analysis. Notably, Oxford Nanopore Technologies (ONT) based sequencing offers the possibility to directly sequence native RNAs without the need for prior cDNA synthesis and PCR amplification. Oxford Nanopore's new long-read cDNA sequencing solutions allow researchers to sequence millions of whole transcripts end-to-end in single reads. This enables users to simultaneously perform accurate isoform-level transcriptome characterization, and differential gene expression profiling.
Fig1. Oxford Nanopore library construction method
Features
High Accuracy
Quantitative Analysis
High Specific
Heterogeneity Analysis
A key advantage of ONT RNA-seq is that multiple features can be addressed simultaneously with high accuracy
All types of ONT transcriptome sequencing libraries are quantifiable
This method can increase the specificity of the comparison between reads and reference genomes
Since nanopore sequencing is a bona fide single-molecule method, molecular heterogeneity at the transcriptome level can be analyzed
Project Workflow
1. Sample Preparation
2. Library Preparation
3. Sequencing
4. Data Analysis & Delivery
Data analysis
Protein interaction analysis
Function enrichment analysis
Detection of fusion transcripts
Identification of transcript boundaries
Gene function and transcript structure annotation
Quantitative expression analysis of transcriptional isoforms
Identification and trimming of full-length sequenced transcripts
Fig1. Oxford Nanopore library construction method
