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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 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 in an environmentally friendly temperature-controlled shipping box.
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.
