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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:
Feature | Property |
Preparation time | 270 minutes |
Input requirement | 100 ng poly-A+ RNA |
Reverse transcription required | Yes |
PCR required | No |
Read length | Enriched for ful-length cDNA during second strand synthesis |
Read type produced | 1D |
Expected data* | ••• |
Typical number of reads | 5-10 million full-length per flow cell on MinION/GirdION; 15-30 million per flow cell on PromethION |
Associated protocols | Direct cDNA Sequencing |
Multiplexing options |
Native Barcoding Expansion Pack 1-12 (EXP-NBD104) Native Barcoding Expansion Pack 13-24 (EXP-NBD114) Flow Cell Wash Kit |
Pack size | 6 reactions |
Stability |
Shipped at -20°C Long-term storage -20°C |
Description
The Direct cDNA Sequencing Kit is used to prepare cDNA for nanopore sequencing without using PCR. The kit recommends an input of 100 ng poly-A+RNA.
Taking full-length poly-A+ RNA, complementary strand synthesis and strand switching are performed using kit-supplied oligonucleotides. The RNA template is then digested, and the second (complementary) strand is synthesised. Sequencing adapters supplied in the Direct cDNA Sequencing Kit are then ligated onto the dscDNA.
Further considerations:
A Native Barcoding Expansion pack (EXP-NBD104) can be used to multiplex up to 24 different RNA samples into a single flow cell.
Oxford Nanopore Technologies deem the useful life of the product to be 3 months from receipt by the customer.