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This kit is recommended for users who:
- Want to target specific regions of 5-20 kb to high coverage (>100x) on a single MinION Mk 1B flow cell without PCR.
- Want to sequence a larger, contiguous Region of Interest (ROI), using up to 100 target sites, in a single assay*
- Have 1-10 µg of available high quality gDNA
- Are interested in methylation patterns or other modified bases.
- Have gene targets that are highly repetitive, or wish to evaluate the number of repeats in an expansion, where traditional amplification method or sequencing-by-synthesis methods could yield a biased result
- Want to sequence long gene targets in a single pass that are not amenable to long-range PCR (>30 kb)
* Note: This is known as ‘tiling’ an ROI.
Feature | Property |
Preparation time | 110 minutes |
Input requirement | 1–10 µg of double-stranded DNA (5 µg recommended) |
PCR Required | No |
Fragmentation | Not required |
Read length | Equal to fragment length |
Read type produced | 1D |
Typical throughput* | ••• |
Associated protocols/documentation |
Cas9 Sequencing Kit (SQK-CS9109) Targeted, amplification-free DNA sequencing using CRISPR/Cas |
Associated analysis tools |
Cas9 targeted sequencing tutorial in EPI2ME Labs Evaluation of read-mapping characteristics from a Cas9 targeted sequencing experiment tutorial |
Multiplexing options | Please contact Technical Support to discuss multiplexing |
Pack size | 6 reactions |
Further Considerations:
Cas9 Targeted sequencing is a fast, flexible method for sequencing targets of interest without amplification. SQK-CS9109 offers users our protocol with the convenience and cost reduction of a kit.
Best coverage is achieved when using the CRISPR/Cas system to target a 5–20 kb region cutting twice at either end. For applications where only one end of the region is known, a single cut and read-out approach is possible.
For users that wish to target regions longer than 20 kb, we suggest a tiling approach and/or ensuring as highly intact starting DNA material as possible. It will not be possible to span larger regions without intact starting gDNA.
We strongly recommend starting with multiple probes and working out which approach works best for your experimental aims, DNA quality etc. Detailed recommendations can be found in the Targeted, amplification-free DNA sequencing using CRISPR/Cas document.
Licenses
Shipping and logistics:
Flow cells and kits are shipped together in a temperature-controlled shipping box with gel ice packs. Ice packs are stored at 2-8° C, and -20° C ice packs are added to the top of the box (but not in direct contact with product) to maintain the integrity of the shipment.
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.