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
|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|
RNA sample quantity ≥ 25 µg.
OD260/230 ≥ 2.0, 2.0 ≤ OD260/280 ≤ 2.2, 28S:18S ≥ 1.0, RIN ≥ 8.
RNA must be treated with DNA enzymes to ensure that it does not contain DNA.
Please make sure that the RNA is not significantly degraded.
Deliverable: FastQ, BAM, QC report, alternative splicing identification, enrichment analysis, KEGG/ GO analysis, custom bioinformatics analysis.