Direct RNA
Sequence RNA molecules directly and preserve base modifications
Up to 1 million reads
PCR cDNA
Optimised for throughput
Up to 12+ million reads
Direct cDNA
No PCR bias
Up to 10 million reads
| | Direct RNA Sequencing Kit | PCR cDNA Sequencing Kit | Direct cDNA Sequencing Kit |
|---|
| Preparation time |
105 mins |
165 mins |
270 mins |
| Input requirement |
500 ng poly-A+ RNA |
1 ng poly-A+ or 50 ng total RNA |
100 ng poly-A+ RNA |
| RT Required |
Optional |
Yes |
Yes |
| PCR Required |
No |
Yes |
No |
| Read length |
Equal to RNA length |
Enriched for full-length cDNA |
Enriched for full-length cDNA |
| Typical throughput |
One star
Less than 1 Gb in 6 hours
1-4 Gb in 48 hours |
Three stars
2-3 Gb in 6 hours
8+ Gb in 48 hours |
Two stars
1-2 Gb in 6 hours
4-8 Gb in 48 hours |
| Typical number of reads |
1 million |
7 - 12+ million |
5 - 10 million |
| Multiplexing options |
In development |
PCR Barcoding Kit |
Native Barcoding Expansion 1-12 and
Native Barcoding Expansion 13-24 |
Strand-switching protocols for preparation of 1D and 2D cDNA libraries
The Ligation Sequencing Kit contains a protocol for 1D cDNA sequencing.
Following first-strand synthesis, reverse transcriptases tend to add 1–3 non-templated Cs to the 3’ end of the cDNA strand, and by including an additional primer in the reaction which anneals to the non- templated Cs, we can make the RT enzyme switch templates, to extend opposite this primer. This incorporates a PCR-priming sequence. Following PCR, sequencing adapters are attached.
Direct RNA offers native strand sequencing, quick prep, full-length transcripts
Nanopores are capable of analysing RNA as well as DNA, but in contrast to other sequencing technologies, nanopores are the only sequencing technology which can sequence RNA directly, rather than sequencing the products of reverse transcription and PCR reactions. There are several ways to prepare RNA strands for sequencing. Fig. 1 shows one method, where a cDNA strand is synthesised, but not sequenced. An adapter is attached to the RNA strand. This allows efficient threading of the RNA strand into the nanopore, and is pre-loaded with the motor protein which controls the speed of translocation of the strand.
cDNA strand switching workflow
Direct RNA workflow
Welcome
Welcome to the Oxford Nanopore US Dollar store.
If you would like to see prices in another currency, please switch here.
If you have an Oxford Nanopore account, please log in.
