This kit is recommended for users who:
- Want to optimise their sequencing experiment for output
- Wish to sequence 2-3 samples for Whole Genome Sequencing (WGS)
- Need a PCR-free method of multiplexing to preserve additional information, such as base modifications
- Require control over read length
- Would like to utilise upstream processes, such as size selection or whole genome amplification
- Would like to process multiple samples simultaneously, either with a multichannel pipette or a liquid handling robot
This is an Early Access product
For more information about our Early Access programmes, please see this article on product release phases.
Please note that to use this kit, you will need to purchase 3rd party reagents: see the "3rd Party Materials" tab for more detail.
The Multiplex Ligation Sequencing Kit XL is a standalone kit providing 96 unique barcodes to enable PCR-free multiplexing of dsDNA samples such as gDNA and amplicons. This kit provides an easier workflow to enable Whole Genome Sequencing (WGS) by enabling low-plex sequencing of 2-3 samples, allowing users to identify regions more common and others more prone to mutations.
The library preparation is similar to the Ligation Sequencing Kit protocol; DNA ends are repaired and dA-tailed using the NEBNext End Repair/dA-tailing module, and then a unique dT-tailed barcode adapter is ligated on the dA-tailed template. Barcoded samples are then pooled together. Each barcode adapter also has a cohesive end and this is used as a hook to ligate to the supplied sequencing adapters.
We recommend starting with 1 μg gDNA per sample and our manual protocol recommends sequencing two samples per flow cell enabling the sequencing of 96 samples across 48 flow cells. There is also an automated protocol on the Hamilton NGS STAR 96 (NGS STAR with Multi-Probe Head 96) to sequence either two samples per flow cell or three samples across two flow cells which is fully supported by Oxford Nanopore Technologies.
The Multiplex Ligation Sequencing Kit XL (Kit 11) includes the Adapter Mix II T (AMII T), with two key features: Firstly, the adapter is higher capture, enabling lower flow cell loading amounts. Secondly, the adapter contains the fuel fix technology, enabling users to run long experiments without the need for fuel addition during the run.
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.
The library preparation method is similar to the Ligation Sequencing Kit protocol; DNA ends are repaired and dA-tailed using the NEBNext End Repair/dA-tailing module, and then a unique dT-tailed barcode adapter is ligated on the dA-tailed template. Barcoded samples are then pooled together. Each barcode adapter also has a cohesive end and this is used as a hook to ligate to the supplied sequencing adapters.
What's in the box
The Multiplex Ligation Sequencing Kit contains enough reagents for 48 reactions.
|Name||Acronym||Cap colour||Number of vials||Fill volume per vial (µl)|
|Adapter Mix II T||AMII T||Green||1||320|
|Sequencing Buffer II||SBII||Red||4||1,500|
|Loading Beads II||LBII||Pink||2||1,500|
|Loading Solution||LS||White cap, pink sticker||2||1,500|
|Elution Buffer||EB||15 ml bottle||1||10,000|
|Long Fragment Buffer||LFB||30 ml bottle||3||15,000|
|Flush Buffer XL||FB||30 ml bottle||4||15,500|
|Flush Tether||FLT||White cap, purple sticker||1||1,600|
|Native Barcodes||NB01-96||N/a||1 plate||8 µl per well|
The Multiplex Ligation Sequencing Kit can be used together with:
- Flow Cell Wash Kit (EXP-WSH004)
- Sequencing Auxiliary Vials (EXP-AUX002)