Nanopore direct RNA sequencing is a third generation sequencing (TGS)-based method to sequence the whole transcriptome of cells or tissues. We provide nanopore direct RNA sequencing to help you resolve structural variants and repetitive regions, enhance metagenomic identification of closely related species, sequence entire microbes in single reads-in real-time.
Overview
Nanopore direct RNA sequencing is one kind of long-read sequencing, emerged in 2011, provided by Oxford Nanopore Technologies (ONT). Nanopore direct RNA sequencing provides a direct, real-time sequencing of long RNA fragments. Nanopore direct RNA sequencing using nanopore inserted in an electrical resistant membrane, there are 2048 nanopores on the membrane. As the RNA fragments pass through the nanopores, the bound polymerase or enzyme can attach to the RNA, the electrical current change was measured, indicating a specific RNA sequence. Nanopore direct RNA sequencing allows ultra-long reads, the longest fragment is close to 1 million bp. Advantages of Nanopore direct RNA sequencing include: 1) direct, real-time analysis; 2) allowing ultra-long kilobases in a continuous read; 3) pocket-sized sequencers are portable with low cost.
We provide Nanopore direct RNA sequencing services for identifying new genes, transcripts, and alternative splicing events, resolving structural variants and repetitive regions, enhancing metagenomic identification of closely related species, sequencing entire microbes in single reads-in real-time. Nanopore direct RNA sequencing also performs rapid pathogen detection and strain characterization, for example, it can be used to screen Ebola and Lassa virus outbreak in Africa.
Features
High throughput
Real-time Analysis
High Resolution
Multiple Applications
This method allow long reads for accurate analysis with a short sequencing time.
Direct, real-time sequencing of long RNA fragments.
Discovery of more cellular differences based on high resolution analysis.
Identify new genes, transcripts and alternative splicing events.
Project Workflow
1. Sample Preparation
RNA purification; quality assessment and quantification.
2. Library Preparation
DNA library preparation.
3. Sequencing
Real-time analysis, takes as little as 10 minutes to perform.
4. Data Analysis
Visualize and preprocess results, and perform custom bioinformatics analysis.
Data Analysis Workflow
In-depth data analysis:
Identification of new genes
Identification of new transcripts
Alternative splicing events comparation between species
Metagenomic identification of closely related species
Sequencing entire microbes in single reads-in real time
Pathogen detection and strain characterization
Sample Requirements
RNA sample (quantity ≥ 100 pg), 1.8 ≤ OD260/280 ≤ 2.2, OD260/230≥2.0, RIN ≥ 6.5, 28S:18S≥1.0. Please make sure that RNA is not significantly degraded.
Sample storage: The sample should be stored at -80°C. Avoid repeated freezing and thawing.
Shipping Method: When shipping the sample, it is stored in a 1.5 mL Eppendorf tube, sealed with a sealing film. Shipments are generally recommended to contain 5-10 pounds of dry ice per 24 hours.
