Sequencing RNA with MinION
Coming next: Direct RNA - the analysis of native RNA
Nanopore sequencing on the MinION is now established as a powerful tool for the analysis of RNA
Researchers can choose between direct RNA analysis – now available for the first time - or cDNA analysis
Direct RNA analysis kits were shipped to early users in late 2016 for the first time.
How is the MinION being used in RNA research?
Available now for sequencing cDNAs
- Gene expression studies
- Long reads to study splice variants and fusion proteins
- Identification of sense and antisense transcripts
- Field-based rapid characterisation of RNA viruses and related epidemiology
Coming next: direct analysis of RNA
- RNA modifications
- PCR-free protocols
- Increased accuracy compared to using reverse transcriptases
RNA analysis is used in a wide variety of experiments including the analysis of full-length transcripts to elucidate structural variations and the rapid identification of viruses.
Nanopore sequencing on the MinION gives you:
Full length transcripts from long reads
Nanopores sequence the whole of the nucleic acid fragment presented to them. For cDNA this means full-length transcripts can be sequenced, simplifying the analysis of spice variants, isoforms and fusion transcripts. Methods are available for poly A+ and random priming cDNA libraries for these transcriptomics studies.Publications on full length transcripts
Nanopore sequencing data starts streaming immediately, rather than being delivered in bulk at the end of a 'run'. Real-time data streaming allows rapid identification of sequences within the sample. Depending on the size of the molecule being sequenced, it is possible that the full cDNA transcript will be sequenced in a single read, simplifying mapping to the genome.Publications on real-time analysis
Nanopore sequencing analyses native nucleic acid
Nanopore sequencing of DNA allows users to sequence without any amplification, which can introduce bias. This has given the opportunity to explore nucleotide modifications by examining and interpreting the nanopore signal produced in the presence of methylation patterns. Released protocols for RNA sequencing on the MinION use traditional approaches which involve generation of cDNA libraries and these can be explored now by starting to use the MinION. But we have shown that RNA can be sequenced directly, which can potentially provide new information on modifications and other properties previously hidden by amplification or reverse transcription. Register your interest at the top of this page to be kept informed.Publications on native nucleic acid research
The MinION weighs under 100g and can fit in a pocket. It is powered by the USB port on a laptop and is uniquely transportable into the field. With library preparation possible using only a minimum of equipment, field-based analysis labs have been setup in remote destinations. Oxford Nanopore is also preparing to release automated sample and library preparation devices to simplify the workflow even further.Publications on field-based use
How are others using the MinION to analyse RNA?
View these publications and movies to find out more.
|24/03/2017||Nanopores allow direct sequencing of full-length RNA strands and modified RNA nucleotides|
|25/11/2016||The Oxford Nanopore MinION: delivery of nanopore sequencing to the genomics community||BioMed Central - Genome Biology|
|12/08/2016||Highly parallel direct RNA sequencing on an array of nanopores||bioRxiv|
|03/08/2016||On the study of microbial transcriptomes using second- and third-generation sequencing technologies||Pub Med|
|26/05/2016||Progress at UC Santa Cruz: Long DNA fragments, tRNA and Modified Bases||London Calling event video|
Progress at UC Santa Cruz: Long DNA fragments, tRNA and Modified Bases
Is being able to analysis native RNA important to your research?
Why not get started with MinION now to become familiar with nanopore sequencing? The MinION starter pack is available for a fee of $1000. The starter pack includes everything that is needed to explore the technology and investigate how the potential of real-time analysis, long reads and being able to use the pocket-sized MinION anywhere could help to answer the biological question. It will take you from never having sequenced using nanopore technology to understanding the capability of the technology and its applicability to your own experimental requirements.
How do I get started?
You can start using MinION straight away.
For a $1,000 fee you receive a MinION and starter pack of flow cells and kits. You also get access to a thriving online community of MinION users.