11th September 2025
- In 'Overview of the protocol', in the Table under 'Library preparation', the timing for priming and loading a flow cell was increased from 5 min to 10 min.
- 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.
- In section 6, 'DNA repair and end-prep', 'Check your flow cell' and section 8, 'Priming and loading the flow cell', 'Complete a flow cell check', the links for the flow cell check in the MinKNOW protocol were replaced with links for the flow cell check document.
- In section 6, 'DNA repair and end-prep', 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 flow cell', the instruction for removing the flow cell out of the fridge was moved to the top of this section.
- In section 12, 'Flow cell reuse and returns', text was edited to advise users not to reuse their flow cells when using this protocol.
- In 'Troubleshooting', section 14, the video links for priming and loading a MinION Flow Cell were replaced with links for the PromethION Flow Cell.
8/12/2025
GDH_9174_v114_revS_12Aug2025
English
PromethION
12 Aug 2025
- Corrected TE buffer formulation to (10 mM Tris-HCl, 1 mM EDTA, pH 8.0) in the "Size selection of gDNA using SFE Buffer" section of the protocol. Previously said 0.1 mM EDTA.
1/22/2025
GDH_9174_v114_revR_22Jan2025
English
PromethION
22 January 2025
- Document title change
- Extraction: Minor adjustments made to mitigate steps where errors may occur.
- Size selection using SFE: Updated method in-line with latest recommendations.
- Megaruptor shearing: Added more context for handling the device effectively.
- Flow cell wash and reload: updated data images and included image for sequencing read length profile.
8/22/2024
GDH_9174_v114_revQ_10Nov2022
English
PromethION
21st August 2024
- Updated sample preparation instructions.
- Updated sample input recommendation into library preparation.
- 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.
- Data acquisition and basecalling instructions updated.
- Downstream analysis section updated.
31 July 2024
- 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
30 April 2024
- Added Megaruptor3 shearing step at the end of Sample extraction to increase sequencing output.
- Updated 'Downstream analysis' instructions.
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
17th July 2023
- Updated "washing and reloading a PromethION Flow Cell" to include recommendation to remove waste after 1 hour incubation with nuclease
10th July 2023
- Updated "Data acquisition and basecalling" to use the most recent MinKNOW GUI version in screenshots
- Updated "Downstream analysis" with the most recent recommendations from EPI2ME labs to ensure successful sequencing and data analysis
- Updated recommendation in the "Downstream analysis" to use modkit not modbam2bed has been depricated.
10th July 2023
- Updated "Data acquisition and basecalling" to use the most recent MinKNOW GUI version in screenshots
- Updated "Downstream analysis" with the most recent recommendations from EPI2ME labs to ensure successful sequencing and data analysis
4th July 2023
- Reformated the "Washing and reloading a PromethION Flow Cell" to be consistent with our other flow cell washing protocols and included an extra recommendation to remove waste buffer between priming mix flushes after using the flow cell wash kit.
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