21st August 2024
- NEBNext® Companion Module v2 for Oxford Nanopore Technologies® Ligation Sequencing (NEB, E7672S or E7672L) is now the recommended reagent module.
- Updated the Repair and End-prep method in-line with new recommended reagents.
- Updated the Adapter ligation step with new recommended method.
- Updated downstream analysis section: recommended software versions.
21st August 2024
- NEBNext® Companion Module v2 for Oxford Nanopore Technologies® Ligation Sequencing (NEB, E7672S or E7672L) is now the recommended reagent module.
- Updated the Repair and End-prep method in-line with new recommended reagents.
- Updated the Adapter ligation step with new recommended method.
- Updated downstream analysis section: recommended software versions.
31 July 2024
- Updated introduction format and inclusion of reference to the RRMS multiplexing protocol.
- Expanded sample preparation section to now include full instructions for DNA extraction and fragmentation.
- Changed reagent order addition in Library preparation/Adapter ligation and clean-up; Ligation Adapter (LA) added before Ligation Buffer (LB) and NEBNext Quick T4 Ligase.
- Updated library preparation method: final elute of adapted DNA library now performed in 38 μl. Allows for users to take split the final elute in three and load directly on to the flow cell (no need to dilute).
- Added instructions on washing and reloading the Flow Cell.
- Updated sequencing run setting instructions to condensed format.
- Re-vamped downstream analysis with new instructions and method.
07 May 2024
- Downstream analysis instructions updated: <br> • human_variation removed as no longer an option <br> • syntax updates for ease and clarity of use <br> • instructions simplified and streamlined <br> • inclusion of additional modkit resource links for more information
DRAFT
20th September 2023
- Updated flow cell loading instructions to include light shield installation
- Updated protocol to include instructions for mouse samples as well as the original method for human samples.
- Updated software recommendations from Guppy to now use Dorado.
- Updated sequencing and library reload times for optimal pore occupancy and flow cell health.
- Updated downstream analysis instructions to Dorado commands and to include information on nextflow pipelines for wf-human-variation.
- Updated downstream analysis instructions to use modkit bioinformatics tool.
1st August 2023
- Kit contents layout has been updated
- In priming and loading the flow cell, the flow cell priming mix instructions have been updated
- Kit updated to released phase and guppy_basecaller parameter updated in "Downstream analysis"
March 2023
- Updated "Data acquisition and basecalling" to correctly reflect basecalling being carried out post-sequencing in the Dpwnstream analysis section of the protocol.
- Added "Ending the experiment" page to the "Sequencing and data analysis" section of the protocol.