Direct cDNA Sequencing Kit
This kit is highly recommended for users who:
- would like to identify and quantify full-length transcripts
- are interested in differential gene expression
- want to characterise and quantify isoforms, splice variants and fusion transcripts
- wish to avoid bias and artefacts introduced through PCR
The Direct cDNA Sequencing Kit features:
Key features:
Prep time | 270 minutes |
Input amount | 100 ng poly-A+ RNA |
Read length | Enriched for full-length cDNA |
Typical throughput | 2-3+ Gb in 6 hours, 8+ Gb in 48 hours per flow cell on MinION/GridION; 10-15+ Gb in 6 hours, 40+ Gb in 48 hours per flow cell on PromethION |
Workflow
Taking full-length messenger RNA, the complementary strand is synthesised using a kit-supplied oligo adapter. The RNA is then degraded, and the second (complementary) strand synthesised. Adapters supplied in the Direct cDNA Sequencing Kit are then ligated onto the cDNA, which introduce the components needed for the cDNA strands to enter the pore. One strand of the duplex is sequenced at a time, producing 1D reads.
