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Ligation sequencing amplicons - Native Barcoding Kit 24 V14 (SQK-NBD114.24) (NBA_9168_v114_revT_02Oct2025)

DateVersionLanguageDeviceChanges
10/8/2025NBA_9168_v114_revT_02Oct2025EnglishGridION
2nd October 2025 - In 'Overview of the protocol', 'Compatibility of this protocol', MinION Mk1B and Mk1C were removed, along with their links for IT requirements as these device documents have moved to legacy. This also includes removal of links to their user manuals in 'Data acquisition and basecalling'. - In section 4, 'Native barcode ligation', an important note was included to only use the barcode wells once and in steps 9, 10 and 12, text was removed for the clear cap EDTA (a reagent in the previous version of the SQK-NBD114.24 kit) as it is no longer applicable. And in step 15, the text was updated to resuspend the beads in SFB by flicking. - In section 5, 'Adapter ligation and clean-up', 'Check your flow cell' and section 6, '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, '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. And the addition of BSA to the flow cell priming mix was changed from a 'recommended' to a 'required' instruction. - In 'Troubleshooting', video links for the PromethION Flow Cell loading were included alongside the MinION Flow Cell loading.
10/8/2025NBA_9168_v114_revT_02Oct2025EnglishMinION
2nd October 2025 - In 'Overview of the protocol', 'Compatibility of this protocol', MinION Mk1B and Mk1C were removed, along with their links for IT requirements as these device documents have moved to legacy. This also includes removal of links to their user manuals in 'Data acquisition and basecalling'. - In section 4, 'Native barcode ligation', an important note was included to only use the barcode wells once and in steps 9, 10 and 12, text was removed for the clear cap EDTA (a reagent in the previous version of the SQK-NBD114.24 kit) as it is no longer applicable. And in step 15, the text was updated to resuspend the beads in SFB by flicking. - In section 5, 'Adapter ligation and clean-up', 'Check your flow cell' and section 6, '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, '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. And the addition of BSA to the flow cell priming mix was changed from a 'recommended' to a 'required' instruction. - In 'Troubleshooting', video links for the PromethION Flow Cell loading were included alongside the MinION Flow Cell loading.
10/8/2025NBA_9168_v114_revT_02Oct2025EnglishPromethION
2nd October 2025 - In 'Overview of the protocol', 'Compatibility of this protocol', MinION Mk1B and Mk1C were removed, along with their links for IT requirements as these device documents have moved to legacy. This also includes removal of links to their user manuals in 'Data acquisition and basecalling'. - In section 4, 'Native barcode ligation', an important note was included to only use the barcode wells once and in steps 9, 10 and 12, text was removed for the clear cap EDTA (a reagent in the previous version of the SQK-NBD114.24 kit) as it is no longer applicable. And in step 15, the text was updated to resuspend the beads in SFB by flicking. - In section 5, 'Adapter ligation and clean-up', 'Check your flow cell' and section 6, '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, '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. And the addition of BSA to the flow cell priming mix was changed from a 'recommended' to a 'required' instruction. - In 'Troubleshooting', video links for the PromethION Flow Cell loading were included alongside the MinION Flow Cell loading.
10/3/2025NBA_9168_v114_revT_02Oct2025EnglishGridION
2nd October 2025 - In 'Overview of the protocol', 'Compatibility of this protocol', MinION Mk1B and Mk1C were removed, along with their links for IT requirements as these device documents have moved to legacy. This also includes removal of links to their user manuals in 'Data acquisition and basecalling'. - In section 4, 'Native barcode ligation', an important note was included to only use the barcode wells once and in steps 9, 10 and 12, text was removed for the clear cap EDTA (a reagent in the previous version of the SQK-NBD114.24 kit) as it is no longer applicable. And in step 15, the text was updated to resuspend the beads in SFB by flicking. - In section 5, 'Adapter ligation and clean-up', 'Check your flow cell' and section 6, '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, '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. And the addition of BSA to the flow cell priming mix was changed from a 'recommended' to a 'required' instruction. - In 'Troubleshooting', video links for the PromethION Flow Cell loading were included alongside the MinION Flow Cell loading.
10/3/2025NBA_9168_v114_revT_02Oct2025EnglishMinION
2nd October 2025 - In 'Overview of the protocol', 'Compatibility of this protocol', MinION Mk1B and Mk1C were removed, along with their links for IT requirements as these device documents have moved to legacy. This also includes removal of links to their user manuals in 'Data acquisition and basecalling'. - In section 4, 'Native barcode ligation', an important note was included to only use the barcode wells once and in steps 9, 10 and 12, text was removed for the clear cap EDTA (a reagent in the previous version of the SQK-NBD114.24 kit) as it is no longer applicable. And in step 15, the text was updated to resuspend the beads in SFB by flicking. - In section 5, 'Adapter ligation and clean-up', 'Check your flow cell' and section 6, '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, '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. And the addition of BSA to the flow cell priming mix was changed from a 'recommended' to a 'required' instruction. - In 'Troubleshooting', video links for the PromethION Flow Cell loading were included alongside the MinION Flow Cell loading.
10/3/2025NBA_9168_v114_revT_02Oct2025EnglishPromethION
2nd October 2025 - In 'Overview of the protocol', 'Compatibility of this protocol', MinION Mk1B and Mk1C were removed, along with their links for IT requirements as these device documents have moved to legacy. This also includes removal of links to their user manuals in 'Data acquisition and basecalling'. - In section 4, 'Native barcode ligation', an important note was included to only use the barcode wells once and in steps 9, 10 and 12, text was removed for the clear cap EDTA (a reagent in the previous version of the SQK-NBD114.24 kit) as it is no longer applicable. And in step 15, the text was updated to resuspend the beads in SFB by flicking. - In section 5, 'Adapter ligation and clean-up', 'Check your flow cell' and section 6, '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, '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. And the addition of BSA to the flow cell priming mix was changed from a 'recommended' to a 'required' instruction. - In 'Troubleshooting', video links for the PromethION Flow Cell loading were included alongside the MinION Flow Cell loading.
10/2/2025NBA_9168_v114_revT_02Oct2025EnglishGridION
2nd October 2025 - In 'Overview of the protocol', 'Compatibility of this protocol', MinION Mk1B and Mk1C were removed, along with their links for IT requirements as these device documents have moved to legacy. This also includes removal of links to their user user manuals in 'Data acquisition and basecalling'. - In section 4, 'Native barcode ligation', an important note was included to only use the barcode wells once and in steps 9, 10 and 12, text was removed for the clear cap EDTA (a reagent in the previous version of the SQK-NBD114,24 kit) as it is no longer applicable. And in step 15, the text was updated to resuspend the beads in SFB by flicking. - In section 5, 'Adapter ligation and clean-up', 'Check your flow cell' and section 6, '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, '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. And the addition of BSA to the flow cell priming mix was changed from a 'recommended' to a 'required' instruction. - In 'Troubleshooting', video links for the PromethION Flow Cell loading were included alongside the MinION Flow Cell loading.
10/2/2025NBA_9168_v114_revT_02Oct2025EnglishMinION
2nd October 2025 - In 'Overview of the protocol', 'Compatibility of this protocol', MinION Mk1B and Mk1C were removed, along with their links for IT requirements as these device documents have moved to legacy. This also includes removal of links to their user user manuals in 'Data acquisition and basecalling'. - In section 4, 'Native barcode ligation', an important note was included to only use the barcode wells once and in steps 9, 10 and 12, text was removed for the clear cap EDTA (a reagent in the previous version of the SQK-NBD114,24 kit) as it is no longer applicable. And in step 15, the text was updated to resuspend the beads in SFB by flicking. - In section 5, 'Adapter ligation and clean-up', 'Check your flow cell' and section 6, '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, '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. And the addition of BSA to the flow cell priming mix was changed from a 'recommended' to a 'required' instruction. - In 'Troubleshooting', video links for the PromethION Flow Cell loading were included alongside the MinION Flow Cell loading.
10/2/2025NBA_9168_v114_revT_02Oct2025EnglishPromethION
2nd October 2025 - In 'Overview of the protocol', 'Compatibility of this protocol', MinION Mk1B and Mk1C were removed, along with their links for IT requirements as these device documents have moved to legacy. This also includes removal of links to their user user manuals in 'Data acquisition and basecalling'. - In section 4, 'Native barcode ligation', an important note was included to only use the barcode wells once and in steps 9, 10 and 12, text was removed for the clear cap EDTA (a reagent in the previous version of the SQK-NBD114,24 kit) as it is no longer applicable. And in step 15, the text was updated to resuspend the beads in SFB by flicking. - In section 5, 'Adapter ligation and clean-up', 'Check your flow cell' and section 6, '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, '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. And the addition of BSA to the flow cell priming mix was changed from a 'recommended' to a 'required' instruction. - In 'Troubleshooting', video links for the PromethION Flow Cell loading were included alongside the MinION Flow Cell loading.
7/2/2025NBA_9168_v114_revS_02Jul2025EnglishGridION
02 July 2025 - Updated protocol introduction section to condensed format. - Increased the timing for priming and loading the flow cell from 5 min to 10 min in Library preparation Table. - Added P2i as a compatible device in 'Compatibility of this protocol'. - Updated information for new kit format with increased Short Fragment Buffer (SFB) volume. - Updated wash steps in the native barcode ligation section of the protocol to use SFB instead of 80% Ethanol. - In section 6, '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.
7/2/2025NBA_9168_v114_revS_02Jul2025EnglishMinION
02 July 2025 - Updated protocol introduction section to condensed format. - Increased the timing for priming and loading the flow cell from 5 min to 10 min in Library preparation Table. - Added P2i as a compatible device in 'Compatibility of this protocol'. - Updated information for new kit format with increased Short Fragment Buffer (SFB) volume. - Updated wash steps in the native barcode ligation section of the protocol to use SFB instead of 80% Ethanol. - In section 6, '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.
7/2/2025NBA_9168_v114_revS_02Jul2025EnglishPromethION
02 July 2025 - Updated protocol introduction section to condensed format. - Increased the timing for priming and loading the flow cell from 5 min to 10 min in Library preparation Table. - Added P2i as a compatible device in 'Compatibility of this protocol'. - Updated information for new kit format with increased Short Fragment Buffer (SFB) volume. - Updated wash steps in the native barcode ligation section of the protocol to use SFB instead of 80% Ethanol. - In section 6, '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.
1/30/2025NBA_9168_v114_revR_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."
1/30/2025NBA_9168_v114_revR_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."
1/30/2025NBA_9168_v114_revR_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."
12/12/2024NBA_9168_v114_revQ_12Dec2024EnglishGridION
 12 December 2024 - Addition of links to MinION Mk1D IT requirements in "Computer requirements and software" 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".
12/12/2024NBA_9168_v114_revQ_12Dec2024EnglishMinION
 12 December 2024 - Addition of links to MinION Mk1D IT requirements in "Computer requirements and software" 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".
12/12/2024NBA_9168_v114_revQ_12Dec2024EnglishPromethION
 12 December 2024 - Addition of links to MinION Mk1D IT requirements in "Computer requirements and software" 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".
11/15/2024NBA_9168_v114_revP_20Nov2024EnglishGridION
November 2024 - Updated kit content information and image to include new kit format of SQK-NBD114.24 with plated native barcodes.
11/15/2024NBA_9168_v114_revP_20Nov2024EnglishMinION
November 2024 - Updated kit content information and image to include new kit format of SQK-NBD114.24 with plated native barcodes.
11/15/2024NBA_9168_v114_revP_20Nov2024EnglishPromethION
November 2024 - Updated kit content information and image to include new kit format of SQK-NBD114.24 with plated native barcodes.
8/21/2024NBA_9168_v114_revO_15Sep2022EnglishGridION
21st August 2024 - Removed Early access loading now that kit is in "released" phase. - Removed reference to duplex sequencing. - Updated flow cell loading recommendations based off input fragment size.
8/21/2024NBA_9168_v114_revO_15Sep2022EnglishMinION
21st August 2024 - Removed Early access loading now that kit is in "released" phase. - Removed reference to duplex sequencing. - Updated flow cell loading recommendations based off input fragment size.
8/21/2024NBA_9168_v114_revO_15Sep2022EnglishPromethION
21st August 2024 - Removed Early access loading now that kit is in "released" phase. - Removed reference to duplex sequencing. - Updated flow cell loading recommendations based off input fragment size.
August 21st, 2024NBA_9168_v114_revM_15Sep2022EnglishGridion Download 07th March 2024 - Updated library loading recommendations to 35-50 fmols.
August 21st, 2024NBA_9168_v114_revM_15Sep2022EnglishMinion Download 07th March 2024 - Updated library loading recommendations to 35-50 fmols.
August 21st, 2024NBA_9168_v114_revM_15Sep2022EnglishPromethion Download 07th March 2024 - Updated library loading recommendations to 35-50 fmols.
October 4th, 2023NBA_9168_v114_revL_15Sep2022EnglishPromethion Download 2nd October 2023 - Updated "Adapter ligation and clean-up" to remove additional reference of NEBNext® Quick Ligation Reaction Buffer (NEB, B6058)
October 4th, 2023NBA_9168_v114_revL_15Sep2022EnglishGridion Download 2nd October 2023 - Updated "Adapter ligation and clean-up" to remove additional reference of NEBNext® Quick Ligation Reaction Buffer (NEB, B6058)
October 4th, 2023NBA_9168_v114_revL_15Sep2022EnglishMinion Download 2nd October 2023 - Updated "Adapter ligation and clean-up" to remove additional reference of NEBNext® Quick Ligation Reaction Buffer (NEB, B6058)
October 2nd, 2023NBA_9168_v114_revK_15Sep2022EnglishPromethion Download 21st September 2023 - Updated flow cell loading instructions to include light shield installation - Format change in "Native barcode ligation" step to fix downloaded checklist.
October 2nd, 2023NBA_9168_v114_revK_15Sep2022EnglishGridion Download 21st September 2023 - Updated flow cell loading instructions to include light shield installation - Format change in "Native barcode ligation" step to fix downloaded checklist.
October 2nd, 2023NBA_9168_v114_revK_15Sep2022EnglishMinion Download 21st September 2023 - Updated flow cell loading instructions to include light shield installation - Format change in "Native barcode ligation" step to fix downloaded checklist.
September 19th, 2023NBA_9168_v114_revI_15Sep2022EnglishPromethion Download 14th August 2023 - Updated the equipment and consumables with both formats of kits which are being shipped. - Updated the native barcoding ligation step to include instructions for both EDTA concentrations. - Updated flow cell flushing instructions to include instructions for the bottle format kit.
September 19th, 2023NBA_9168_v114_revI_15Sep2022EnglishGridion Download 14th August 2023 - Updated the equipment and consumables with both formats of kits which are being shipped. - Updated the native barcoding ligation step to include instructions for both EDTA concentrations. - Updated flow cell flushing instructions to include instructions for the bottle format kit.
September 19th, 2023NBA_9168_v114_revI_15Sep2022EnglishMinion Download 14th August 2023 - Updated the equipment and consumables with both formats of kits which are being shipped. - Updated the native barcoding ligation step to include instructions for both EDTA concentrations. - Updated flow cell flushing instructions to include instructions for the bottle format kit.
September 14th, 2023NBA_9168_v114_revH_15Sep2022EnglishPromethion Download April 2023 - Added flow cell priming mix volumes to the "Priming and loading" steps
September 14th, 2023NBA_9168_v114_revH_15Sep2022EnglishGridion Download April 2023 - Added flow cell priming mix volumes to the "Priming and loading" steps
September 14th, 2023NBA_9168_v114_revH_15Sep2022EnglishMinion Download April 2023 - Added flow cell priming mix volumes to the "Priming and loading" steps
April 5th, 2023NBA_9168_v114_revG_15Sep2022EnglishPromethion Download April 2023 - Updated the DNA repair end-prep step mastermix tip.
April 5th, 2023NBA_9168_v114_revG_15Sep2022EnglishGridion Download April 2023 - Updated the DNA repair end-prep step mastermix tip.
April 5th, 2023NBA_9168_v114_revG_15Sep2022EnglishMinion Download April 2023 - Updated the DNA repair end-prep step mastermix tip.
March 24th, 2023NBA_9168_v114_revF_15Sep2022EnglishPromethion Download March 2023 - Updated end-prep step to show the tip for making up an end-prep mastermix before combining the reagents. - Corrected the expected volumes for pooled samples and the reflected in the AXP volumes.
March 24th, 2023NBA_9168_v114_revF_15Sep2022EnglishGridion Download March 2023 - Updated end-prep step to show the tip for making up an end-prep mastermix before combining the reagents. - Corrected the expected volumes for pooled samples and the reflected in the AXP volumes.
March 24th, 2023NBA_9168_v114_revF_15Sep2022EnglishMinion Download March 2023 - Updated end-prep step to show the tip for making up an end-prep mastermix before combining the reagents. - Corrected the expected volumes for pooled samples and the reflected in the AXP volumes.
January 6th, 2023NBA_9168_v114_revE_15Sep2022EnglishPromethion Download January 2023 - Changed the "Priming and loading the PrometION flow cell" section to the updated version.
January 6th, 2023NBA_9168_v114_revE_15Sep2022EnglishGridion Download January 2023 - Changed the "Priming and loading the PrometION flow cell" section to the updated version.
January 6th, 2023NBA_9168_v114_revE_15Sep2022EnglishMinion Download January 2023 - Changed the "Priming and loading the PrometION flow cell" section to the updated version.
October 20th, 2022NBA_9168_v114_revD_15Sep2022EnglishPromethion Download 20th October 2022 - EXP-NBA114 information updated with availability
October 20th, 2022NBA_9168_v114_revD_15Sep2022EnglishGridion Download 20th October 2022 - EXP-NBA114 information updated with availability
October 20th, 2022NBA_9168_v114_revD_15Sep2022EnglishMinion Download 20th October 2022 - EXP-NBA114 information updated with availability
October 18th, 2022NBA_9168_v114_revC_15Sep2022EnglishPromethion Download 18th October 2022 - Updated equipment and consumables to remove reference to the NEBNext Companion Module for Oxford Nanopore Technologies Ligation Sequencing
October 18th, 2022NBA_9168_v114_revC_15Sep2022EnglishGridion Download 18th October 2022 - Updated equipment and consumables to remove reference to the NEBNext Companion Module for Oxford Nanopore Technologies Ligation Sequencing
October 18th, 2022NBA_9168_v114_revC_15Sep2022EnglishMinion Download 18th October 2022 - Updated equipment and consumables to remove reference to the NEBNext Companion Module for Oxford Nanopore Technologies Ligation Sequencing
October 4th, 2022NBA_9168_v114_revB_15Sep2022EnglishPromethion Download 4th Oct 2022 - Updated kit 14 info sheet snippet and link
October 4th, 2022NBA_9168_v114_revB_15Sep2022EnglishGridion Download 4th Oct 2022 - Updated kit 14 info sheet snippet and link
October 4th, 2022NBA_9168_v114_revB_15Sep2022EnglishMinion Download 4th Oct 2022 - Updated kit 14 info sheet snippet and link
September 15th, 2022NBA_9168_v114_revA_15Sep2022EnglishPromethion Download -
September 15th, 2022NBA_9168_v114_revA_15Sep2022EnglishGridion Download -
September 15th, 2022NBA_9168_v114_revA_15Sep2022EnglishMinion Download -

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