19th September 2025
- In 'Steps in the sequencing workflow', in the Table for 'Library preparation', the timing for flow cell priming and loading was increased from 5 min to 10 min and the total timing in the workflow image was updated to reflect this change.
- In 'Compatibility of this protocol', GridION was added as a compatible device.
- In 'Equipment and consumables', the image of the LSK114 kit in single-use tube format was removed. And instructions for the single-use tube format were removed from section 8, step 2 and in section 9, step 17, as the kit has been updated to a bottle format.
-In section 6, 'DNA repair and end-prep', instructions to check your flow cell were added at the start of the section. And instructions for use of the previous version of the NEBNext Companion Module for Oxford Nanopore Technologies Ligation Sequencing (NEB, E7180S or E7180L) were removed as the current Companion Module v2 is more efficient.
- -In 'DNA repair and end-prep', step 5, instruction was added to hold the sample(s) at 4°C. And in step 15, instructions were provided on how to resuspend the AMPure XP bead pellet with nuclease-free water.
- In section 8, 'Priming and loading the MinION and GridION Flow Cell', a tip was added to bring the flow cell to room temperature for better visibility of the array. The addition of BSA to the flow cell priming mix was removed as it is not required for adaptive sampling.
- In section 9, 'Washing and reloading a MinION Flow Cell', the addition of BSA to the flow cell priming mix was removed as it is not required for adaptive sampling. And the step to 'Start a pore scan' was removed as this function is not available.
- In 'Troubleshooting', section 14, Tables referring to Guppy were removed as it is no longer applicable.
12/13/2024
RRMS_9180_v114_revK_13Dec2024
English
GridION
13 December 2024
- Addition of MinION Mk1D device for RRMS in "Overview of the protocol".
- Arrow which was previously at the bottom edge of the flow cell in Image 1b "Priming and loading MinION/GridION Flow Cell" was moved to the middle of the flow cell.
- Updated link for flow cell returns in "Flow cell reuse and returns" and removed note for flushing flow cells.
- Updated link for MinION temperature control in "Issues during a sequencing run", "Failed to reach target temperature".
9/3/2024
RRMS_9180_v114_revJ_08Feb2023
English
GridION
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.
