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Human variation sequencing from 10kb extracted cell line samples using SQK-LSK114 (GDH_9173_v114_revR_10Sep2025)


DateVersionLanguageDeviceChanges
9/11/2025GDH_9173_v114_revR_10Sep2025EnglishPromethION
10th 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 and timings in the workflow image were updated. - 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 7, step 2. - In section 5, 'DNA repair and end-prep', 'Check your flow cell' and section 7, '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 5, '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 7, '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 'Troubleshooting', section 12, the videos links for priming and loading a MinION Flow Cell were replaced with links for the PromethION Flow Cell.
1/22/2025GDH_9173_v114_revQ_22Jan2025EnglishPromethION
22 January 2025 - Document title change - gDNA Extraction: method updated
8/23/2024GDH_9173_v114_revP_10Nov2022EnglishPromethION
23st August 2024 - Reverted the Sample preparation section to Human cell line DNA extraction using the QIAGEN Puregene Cell Kit & Shearing DNA for 10 kb input using the Covaris g-TUBE™ - Sample input recommendation into library preparation reverted to 1 ug. 21st August 2024 - Updated workflow overview image. - 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.
November 10th, 2022GDH_9173_v114_revB_10Nov2022English- Download 10th November 2022 Updated introduction to be inline with the online protocol
November 9th, 2022Prom_end_to_end_10kb_v3English- Download -
August 23rd, 2024GDH_9173_v114_revO_10Nov2022English- Download 23st August 2024 - Reverted the Sample preparation section to Human cell line DNA extraction using the QIAGEN Puregene Cell Kit & Shearing DNA for 10 kb input using the Covaris g-TUBE™ - Sample input recommendation into library preparation reverted to 1 ug. 21st August 2024 - Updated workflow overview image. - 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.
August 23rd, 2024GDH_9173_v114_revN_10Nov2022English- Download 21st August 2024 - Updated workflow overview image. - 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.
August 21st, 2024GDH_9173_v114_revL_31Jul2024English- Download 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 - Updated 'Downstream analysis' instructions.
July 31st, 2024GDH_9173_v114_revJ_10Nov2022English- Download 29th Feb 2024 - Updated consumable TE buffer (10 mM Tris-HCl, 1 mM EDTA, pH 8.0) to product code Fisher scientific, 10224683
February 29th, 2024GDH_9173_v114_revI_10Nov2022English- Download 19th September 2023 - Updated flow cell loading instructions to include light shield installation
September 19th, 2023GDH_9173_v114_revH_10Nov2022English- Download 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
August 1st, 2023GDH_9173_v114_revG_10Nov2022English- Download 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.
July 10th, 2023GDH_9173_v114_revF_10Nov2022English- Download 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
May 3rd, 2023GDH_9173_v114_revE_10Nov2022English- Download May 2023 - This kit has moved to released phase.
April 24th, 2023GDH_9173_v114_revD_10Nov2022English- Download April 2023 - Added flow cell priming mix volumes to the "Priming and loading" steps
January 6th, 2023GDH_9173_v114_revC_10Nov2022English- Download January 2023 - Changed the "Priming and loading the PrometION flow cell" section to the updated version.

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