Ligation sequencing DNA XL V14 (SQK-LSK114-XL) (GDX_9163_v114_revU_28May2025)

DateVersionLanguageDeviceChanges
5/28/2025GDX_9163_v114_revU_28May2025EnglishGridION
28th May 2025 - In 'Overview', library preparation time was increased to ~65 min to align with Table and Figure in 'Steps in the sequencing workflow' where priming and loading of flow cell was increased from 5 min to 10 min. - In section 3 'DNA repair and end-prep', step 7, the following was added to all device arms: 'Then cool down to between 4°C and 20°C on the thermal cycler or place the samples on ice.'. - In section 5, 'Priming and loading the MinION and GridION Flow Cell'', an important note was added to the bottom of the section stating that when a flow cell is inserted into the MinION Mk1D and the lid is closed, there will be a small gap around the sides of the device. - For the PromethION branch, the step for removing the PromethION Flow Cell out of the fridge was moved to the beginning of the section on 'Priming and loading the PromethION Flow Cell' to enable the flow cell to reach room temperature while the flow cell priming mix is prepared. And an optional action of performing a flow cell check was included. - In section 6, 'Data acquisition and basecalling', 'To start a sequencing run in MinKNOW', the sequencing kit was specified in point 3, 'Select the Ligation Sequencing Kit XL V14 (SQK-LSK114) on the Kit page'. And the section on 'Duplex basecalling' was removed as it is no longer applicable.
5/28/2025GDX_9163_v114_revU_28May2025EnglishMinION
28th May 2025 - In 'Overview', library preparation time was increased to ~65 min to align with Table and Figure in 'Steps in the sequencing workflow' where priming and loading of flow cell was increased from 5 min to 10 min. - In section 3 'DNA repair and end-prep', step 7, the following was added to all device arms: 'Then cool down to between 4°C and 20°C on the thermal cycler or place the samples on ice.'. - In section 5, 'Priming and loading the MinION and GridION Flow Cell'', an important note was added to the bottom of the section stating that when a flow cell is inserted into the MinION Mk1D and the lid is closed, there will be a small gap around the sides of the device. - For the PromethION branch, the step for removing the PromethION Flow Cell out of the fridge was moved to the beginning of the section on 'Priming and loading the PromethION Flow Cell' to enable the flow cell to reach room temperature while the flow cell priming mix is prepared. And an optional action of performing a flow cell check was included. - In section 6, 'Data acquisition and basecalling', 'To start a sequencing run in MinKNOW', the sequencing kit was specified in point 3, 'Select the Ligation Sequencing Kit XL V14 (SQK-LSK114) on the Kit page'. And the section on 'Duplex basecalling' was removed as it is no longer applicable.
5/28/2025GDX_9163_v114_revU_28May2025EnglishPromethION
28th May 2025 - In 'Overview', library preparation time was increased to ~65 min to align with Table and Figure in 'Steps in the sequencing workflow' where priming and loading of flow cell was increased from 5 min to 10 min. - In section 3 'DNA repair and end-prep', step 7, the following was added to all device arms: 'Then cool down to between 4°C and 20°C on the thermal cycler or place the samples on ice.'. - In section 5, 'Priming and loading the MinION and GridION Flow Cell'', an important note was added to the bottom of the section stating that when a flow cell is inserted into the MinION Mk1D and the lid is closed, there will be a small gap around the sides of the device. - For the PromethION branch, the step for removing the PromethION Flow Cell out of the fridge was moved to the beginning of the section on 'Priming and loading the PromethION Flow Cell' to enable the flow cell to reach room temperature while the flow cell priming mix is prepared. And an optional action of performing a flow cell check was included. - In section 6, 'Data acquisition and basecalling', 'To start a sequencing run in MinKNOW', the sequencing kit was specified in point 3, 'Select the Ligation Sequencing Kit XL V14 (SQK-LSK114) on the Kit page'. And the section on 'Duplex basecalling' was removed as it is no longer applicable.
1/30/2025GDX_9163_v114_revT_30Jan2025EnglishGridION
30th January 2025 In "Adapter ligation and clean-up". the instructions for preparing the LFB or SFB have been changed to: "Thaw either Long Fragment Buffer (LFB) or Short Fragment Buffer (SFB) at room temperature and mix by vortexing. Then spin down and keep at room temperature."
1/30/2025GDX_9163_v114_revT_30Jan2025EnglishMinION
30th January 2025 In "Adapter ligation and clean-up". the instructions for preparing the LFB or SFB have been changed to: "Thaw either Long Fragment Buffer (LFB) or Short Fragment Buffer (SFB) at room temperature and mix by vortexing. Then spin down and keep at room temperature."
1/30/2025GDX_9163_v114_revT_30Jan2025EnglishPromethION
30th January 2025 In "Adapter ligation and clean-up". the instructions for preparing the LFB or SFB have been changed to: "Thaw either Long Fragment Buffer (LFB) or Short Fragment Buffer (SFB) at room temperature and mix by vortexing. Then spin down and keep at room temperature."
12/12/2024GDX_9163_v114_revS_12Dec2024EnglishGridION
12 December 2024 - Addition of links to MinION Mk1D IT requirements in "Compatibility of this protocol" and user manual in "Data acquisition and basecalling", "How to start sequencing". - 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".
12/12/2024GDX_9163_v114_revS_12Dec2024EnglishMinION
12 December 2024 - Addition of links to MinION Mk1D IT requirements in "Compatibility of this protocol" and user manual in "Data acquisition and basecalling", "How to start sequencing". - 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".
12/12/2024GDX_9163_v114_revS_12Dec2024EnglishPromethION
12 December 2024 - Addition of links to MinION Mk1D IT requirements in "Compatibility of this protocol" and user manual in "Data acquisition and basecalling", "How to start sequencing". - 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".
8/21/2024GDX_9163_v114_revR_29Jun2022EnglishGridION
21st August 2024 - Updated protocol introduction to trimmed down format. - Updated sample input recommendations with table for different DNA size. - 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 library loading on flow cell recommendations with table for different DNA size.
8/21/2024GDX_9163_v114_revR_29Jun2022EnglishMinION
21st August 2024 - Updated protocol introduction to trimmed down format. - Updated sample input recommendations with table for different DNA size. - 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 library loading on flow cell recommendations with table for different DNA size.
8/21/2024GDX_9163_v114_revR_29Jun2022EnglishPromethION
21st August 2024 - Updated protocol introduction to trimmed down format. - Updated sample input recommendations with table for different DNA size. - 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 library loading on flow cell recommendations with table for different DNA size.
August 21st, 2024GDX_9163_v114_revQ_31Jul2024EnglishMinion 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 4th October 2023 - Updated the format of the final step in the "Adapter ligation and clean-up" step for the checklist
August 21st, 2024GDX_9163_v114_revQ_31Jul2024EnglishPromethion 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 4th October 2023 - Updated the format of the final step in the "Adapter ligation and clean-up" step for the checklist
August 21st, 2024GDX_9163_v114_revQ_31Jul2024EnglishGridion 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 4th October 2023 - Updated the format of the final step in the "Adapter ligation and clean-up" step for the checklist
October 4th, 2023GDX_9163_v114_revO_29Jun2022EnglishMinion Download 27th September 2023 - Correct error in an equipment and consumable snippet regarding input recommendation for the library preparation
October 4th, 2023GDX_9163_v114_revO_29Jun2022EnglishPromethion Download 27th September 2023 - Correct error in an equipment and consumable snippet regarding input recommendation for the library preparation
October 4th, 2023GDX_9163_v114_revO_29Jun2022EnglishGridion Download 27th September 2023 - Correct error in an equipment and consumable snippet regarding input recommendation for the library preparation
September 27th, 2023GDX_9163_v114_revN_29Jun2022EnglishMinion Download 19th September 2023 - Updated flow cell loading instructions to include light shield installation - Updated flow cell loading recommendations in the "Adapter ligation and clean-up" step. - Update the equipment and consumables section to only be relevant for high molecular weight DNA
September 27th, 2023GDX_9163_v114_revN_29Jun2022EnglishPromethion Download 19th September 2023 - Updated flow cell loading instructions to include light shield installation - Updated flow cell loading recommendations in the "Adapter ligation and clean-up" step. - Update the equipment and consumables section to only be relevant for high molecular weight DNA
September 27th, 2023GDX_9163_v114_revN_29Jun2022EnglishGridion Download 19th September 2023 - Updated flow cell loading instructions to include light shield installation - Updated flow cell loading recommendations in the "Adapter ligation and clean-up" step. - Update the equipment and consumables section to only be relevant for high molecular weight DNA
September 19th, 2023GDX_9163_v114_revM_29Jun2022EnglishMinion Download May 2023: - Updated the elution incubation step in the "DNA and end-prep" step of the protocol to be uniform with the SQK-LSK114 protocols. - This kit has moved to released phase.
September 19th, 2023GDX_9163_v114_revM_29Jun2022EnglishPromethion Download May 2023: - Updated the elution incubation step in the "DNA and end-prep" step of the protocol to be uniform with the SQK-LSK114 protocols. - This kit has moved to released phase.
September 19th, 2023GDX_9163_v114_revM_29Jun2022EnglishGridion Download May 2023: - Updated the elution incubation step in the "DNA and end-prep" step of the protocol to be uniform with the SQK-LSK114 protocols. - This kit has moved to released phase.
April 3rd, 2023GDX_9163_v114_revL_29Jun2022EnglishMinion Download April 2023: - Updated the elution incubation step in the "DNA and end-prep" step of the protocol to be uniform with the SQK-LSK114 protocols.
April 3rd, 2023GDX_9163_v114_revL_29Jun2022EnglishPromethion Download April 2023: - Updated the elution incubation step in the "DNA and end-prep" step of the protocol to be uniform with the SQK-LSK114 protocols.
April 3rd, 2023GDX_9163_v114_revL_29Jun2022EnglishGridion Download April 2023: - Updated the elution incubation step in the "DNA and end-prep" step of the protocol to be uniform with the SQK-LSK114 protocols.
March 15th, 2023GDX_9163_v114_revK_29Jun2022EnglishMinion Download March 2023: - Updated the "Sequencing and data analysis" section to include further details about duplex basecalling
March 15th, 2023GDX_9163_v114_revK_29Jun2022EnglishPromethion Download March 2023: - Updated the "Sequencing and data analysis" section to include further details about duplex basecalling
March 15th, 2023GDX_9163_v114_revK_29Jun2022EnglishGridion Download March 2023: - Updated the "Sequencing and data analysis" section to include further details about duplex basecalling
March 15th, 2023GDX_9163_v114_revJ_29Jun2022EnglishMinion Download January 2023 - Changed the "Priming and loading the PrometION flow cell" section to the updated version.
March 15th, 2023GDX_9163_v114_revJ_29Jun2022EnglishPromethion Download January 2023 - Changed the "Priming and loading the PrometION flow cell" section to the updated version.
March 15th, 2023GDX_9163_v114_revJ_29Jun2022EnglishGridion Download January 2023 - Changed the "Priming and loading the PrometION flow cell" section to the updated version.
October 27th, 2022GDX_9163_v114_revI_29Jun2022EnglishMinion Download 27th October 2022 - Updated protocol to use 80% ethanol rather than 70% ethanol.
October 27th, 2022GDX_9163_v114_revI_29Jun2022EnglishPromethion Download 27th October 2022 - Updated protocol to use 80% ethanol rather than 70% ethanol.
October 27th, 2022GDX_9163_v114_revI_29Jun2022EnglishGridion Download 27th October 2022 - Updated protocol to use 80% ethanol rather than 70% ethanol.
October 4th, 2022GDX_9163_v114_revH_29Jun2022EnglishMinion Download 4th Oct 2022 - Updated kit 14 info sheet snippet and link
October 4th, 2022GDX_9163_v114_revH_29Jun2022EnglishPromethion Download 4th Oct 2022 - Updated kit 14 info sheet snippet and link
October 4th, 2022GDX_9163_v114_revH_29Jun2022EnglishGridion Download 4th Oct 2022 - Updated kit 14 info sheet snippet and link
August 18th, 2022GDX_9163_v114_revG_29Jun2022EnglishMinion Download 18th August 2022 - Updated kit contents image
August 18th, 2022GDX_9163_v114_revG_29Jun2022EnglishPromethion Download 18th August 2022 - Updated kit contents image
August 18th, 2022GDX_9163_v114_revG_29Jun2022EnglishGridion Download 18th August 2022 - Updated kit contents image
August 10th, 2022GDX_9163_v114_revF_29Jun2022EnglishMinion Download 10th August 2022 - Added specific concentration of BSA required for preparing the flow cell priming mix
August 10th, 2022GDX_9163_v114_revF_29Jun2022EnglishPromethion Download 10th August 2022 - Added specific concentration of BSA required for preparing the flow cell priming mix
August 10th, 2022GDX_9163_v114_revF_29Jun2022EnglishGridion Download 10th August 2022 - Added specific concentration of BSA required for preparing the flow cell priming mix
August 10th, 2022GDX_9163_v114_revE_29Jun2022EnglishMinion Download 10th August 2022 Updated phrasing used to add BSA to the flow cell priming mix and the use of ligation buffer
August 10th, 2022GDX_9163_v114_revE_29Jun2022EnglishPromethion Download 10th August 2022 Updated phrasing used to add BSA to the flow cell priming mix and the use of ligation buffer
August 10th, 2022GDX_9163_v114_revE_29Jun2022EnglishGridion Download 10th August 2022 Updated phrasing used to add BSA to the flow cell priming mix and the use of ligation buffer
August 10th, 2022GDX_9163_v114_revD_29Jun2022EnglishMinion Download 9th August 2022 - "Priming and loading the SpotON flow cell" has been updated to include the use of BSA for MinION and GridION flow cells
August 10th, 2022GDX_9163_v114_revD_29Jun2022EnglishPromethion Download 9th August 2022 - "Priming and loading the SpotON flow cell" has been updated to include the use of BSA for MinION and GridION flow cells
August 10th, 2022GDX_9163_v114_revD_29Jun2022EnglishGridion Download 9th August 2022 - "Priming and loading the SpotON flow cell" has been updated to include the use of BSA for MinION and GridION flow cells
August 10th, 2022GDX_9163_v114_revD_29Jun2022EnglishMinion Download 9th August 2022 - "Priming and loading the SpotON flow cell" has been updated to include the use of BSA for MinION and GridION flow cells
August 10th, 2022GDX_9163_v114_revD_29Jun2022EnglishPromethion Download 9th August 2022 - "Priming and loading the SpotON flow cell" has been updated to include the use of BSA for MinION and GridION flow cells
August 10th, 2022GDX_9163_v114_revD_29Jun2022EnglishGridion Download 9th August 2022 - "Priming and loading the SpotON flow cell" has been updated to include the use of BSA for MinION and GridION flow cells
July 21st, 2022GDX_9163_v114_revC_29Jun2022EnglishMinion Download 21st July 2022 - Updated the flow cell loading recommendations for MinION/GridION and PromethION flow cells
July 21st, 2022GDX_9163_v114_revC_29Jun2022EnglishPromethion Download 21st July 2022 - Updated the flow cell loading recommendations for MinION/GridION and PromethION flow cells
July 21st, 2022GDX_9163_v114_revC_29Jun2022EnglishGridion Download 21st July 2022 - Updated the flow cell loading recommendations for MinION/GridION and PromethION flow cells
July 21st, 2022GDX_9163_v114_revB_29Jun2022EnglishMinion Download 21st July 2022 - Updated flow cell loading recommendations for both MinION/GridION and PromethION - Edited protocol to be inclusive of both gDNA and amplicon DNA inputs: - Overview edited to reflect change above. - Included "Category" and "Sample type" to reflect change above - Included snippet in "Equipment and consumables" and "DNA repair and end- prep" to clarify difference in DNA sample inputs. - Have made "Materials" more inclusive to both DNA input types. - All relevant snippets should now account for both sample input types.
July 21st, 2022GDX_9163_v114_revB_29Jun2022EnglishPromethion Download 21st July 2022 - Updated flow cell loading recommendations for both MinION/GridION and PromethION - Edited protocol to be inclusive of both gDNA and amplicon DNA inputs: - Overview edited to reflect change above. - Included "Category" and "Sample type" to reflect change above - Included snippet in "Equipment and consumables" and "DNA repair and end- prep" to clarify difference in DNA sample inputs. - Have made "Materials" more inclusive to both DNA input types. - All relevant snippets should now account for both sample input types.
July 21st, 2022GDX_9163_v114_revB_29Jun2022EnglishGridion Download 21st July 2022 - Updated flow cell loading recommendations for both MinION/GridION and PromethION - Edited protocol to be inclusive of both gDNA and amplicon DNA inputs: - Overview edited to reflect change above. - Included "Category" and "Sample type" to reflect change above - Included snippet in "Equipment and consumables" and "DNA repair and end- prep" to clarify difference in DNA sample inputs. - Have made "Materials" more inclusive to both DNA input types. - All relevant snippets should now account for both sample input types.
June 30th, 2022GDX_9163_v114_revA_29Jun2022EnglishMinion Download 23rd June 2022 -Finalised full protocol for the Kit 14 contents. Just missing image in the Ligation Sequencing Kit XL (SQK-LSK114-XL) contents 20th June 2022 - Update the priming and loading the SpotON flow cells for both MinION and GridION with new diagrams.
June 30th, 2022GDX_9163_v114_revA_29Jun2022EnglishPromethion Download 23rd June 2022 -Finalised full protocol for the Kit 14 contents. Just missing image in the Ligation Sequencing Kit XL (SQK-LSK114-XL) contents 20th June 2022 - Update the priming and loading the SpotON flow cells for both MinION and GridION with new diagrams.
June 30th, 2022GDX_9163_v114_revA_29Jun2022EnglishGridion Download 23rd June 2022 -Finalised full protocol for the Kit 14 contents. Just missing image in the Ligation Sequencing Kit XL (SQK-LSK114-XL) contents 20th June 2022 - Update the priming and loading the SpotON flow cells for both MinION and GridION with new diagrams.
June 29th, 2022Rev1EnglishMinion Download 23rd June 2022 -Finalised full protocol for the Kit 14 contents. Just missing image in the Ligation Sequencing Kit XL (SQK-LSK114-XL) contents 20th June 2022 - Update the priming and loading the SpotON flow cells for both MinION and GridION with new diagrams.
June 29th, 2022Rev1EnglishPromethion Download 23rd June 2022 -Finalised full protocol for the Kit 14 contents. Just missing image in the Ligation Sequencing Kit XL (SQK-LSK114-XL) contents 20th June 2022 - Update the priming and loading the SpotON flow cells for both MinION and GridION with new diagrams.
June 29th, 2022Rev1EnglishGridion Download 23rd June 2022 -Finalised full protocol for the Kit 14 contents. Just missing image in the Ligation Sequencing Kit XL (SQK-LSK114-XL) contents 20th June 2022 - Update the priming and loading the SpotON flow cells for both MinION and GridION with new diagrams.