Information
This kit is highly recommended for users who:
- have a limited amount of input material
- want to optimise their sequencing experiment for throughput
- 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
Please note that to use this kit, you will need to purchase additional 3rd party reagents: see the "3rd Party Materials" tab for more detail.
The PCR-cDNA Sequencing Kit is used to prepare cDNA for nanopore sequencing from an input of as low as 1 ng poly-A+ RNA. Users who do not have poly-A+ enriched RNA can use 50 ng of total RNA but additional optimisation may be required.
Taking full-length poly-A+ RNA, complementary strand synthesis and strand switching are performed using kit-supplied oligonucleotides. dscDNA is then generated by PCR amplification using primers that contain 5’ tags which facilitate the ligase-free attachment of Rapid Sequencing Adapters.
The PCR-cDNA Sequencing Kit features:
| Feature | Property |
|---|
| Preparation time | 165 minutes |
| 165 minutes | 1 ng poly-A+ RNA (or 50 ng total RNA) |
| Reverse transcription required | Yes |
| PCR required | Yes |
| Read length | Enriched for full-length cDNA during PCR |
| Read type produced | 1D |
| Typical throughput | ★★★ |
| Typical number of reads | 7–12+ million full-length mapped readsper flow cell on MinION/GridION; 50+ million mapped reads per flow cell on PromethION |
| Associated protocols | • cDNA-PCR sequencing
• Sequence-specific cDNA-PCR sequencing |
| Multiplexing options | • PCR-cDNA Barcoding Kit
• Flow Cell Wash Kit |
| Pack size | 6 reactions |
| Stability | Shipped at 2–8°C
Long-term storage -20°C
Oxford Nanopore Technologies deem the useful life of the product to be 3 months from receipt by the customer. |
Further considerations
The PCR-cDNA Barcoding Kit (SQK-PCB109) can be used instead of the PCR-cDNA Sequencing Kit to multiplex up to 12 different RNA samples into a single flow cell.
Shipping and logistics:
Flow cells and kits are shipped together at 2–8°C.
Upon receipt, please place the right product in the right long-term storage location.
Products are shipped to customers within the USA and EU Monday to Thursday. Shipments to Canada, Norway, Korea and Japan are expedited Monday to Wednesday; with Australia and New Zealand leaving our warehouses on a Friday. Shipments to the rest of the world are made on Mondays to allow the full working week for packages to arrive.
The delivery charges are calculated when a quote is raised or during checkout. Once an order is made, the delivery ID and delivery information can be tracked in the Store.
Workflow
Taking full-length PolyA+ RNA, complementary strand synthesis and strand switching are performed using kit-supplied oligonucleotides. dscDNA is then generated by PCR amplification using primers that contain 5’ tags which facilitate the ligase-free attachment of Rapid Sequencing Adapters.
Protocols
• cDNA-PCR sequencing
• Sequence-specific cDNA-PCR sequencing
To access the protocols, you will need to register for a Nanopore Community account.
What's in the box
The PCR-cDNA Sequencing Kit contains sufficient reagents to generate six sequencing libraries.
| Name | Acronym | Cap colour | No. of vials | Fill volume per vial (μl) |
|---|
| VN Primer | VNP | Blue | 1 | 200 |
| Strand Switching Primer | SSP | Pink | 1 | 160 |
| cDNA Primer | cPRM | Grey | 1 | 40 |
| Elution Buffer | EB | Black | 2 | 200 |
| Rapid Adapter | RAP | Green | 1 | 10 |
| Sequencing Buffer | SQB | Red | 2 | 300 |
| Sequencing Tether | SQT | Purple | 1 | 10 |
| Loading Beads | LB | Pink | 1 | 360 |
The Flow Cell Priming Kit is also supplied.
![EXP-FLP002]()
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| Name | Acronym | Cap colour | No. of vials | Fill volume per vial (μl) |
|---|
| Flush Buffer | FB | Blue | 6 | 1,170 |
| Flush Tether | FLT | Purple | 1 | 200 |
Compatibility
The PCR-cDNA Sequencing Kit can be used together with:
Kits
- Flow Cell Wash Kit (EXP-WSH004)
Flow cells
- FLO-MINSP6
- FLO-MIN106D
- FLO-PRO002
Devices
- MinION Mk1B
- MinION Mk1C
- GridION
- PromethION
