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Ligation sequencing DNA V14 - dual barcoding (SQK-NBD114.24 with EXP-PBC096) (DBC_9184_v114_revN_02Jul2025)


DateVersionLanguageDeviceChanges
7/2/2025DBC_9184_v114_revN_02Jul2025EnglishGridION
02 July 2025 - Updated overview of the protocol section within the introduction summary to a table format for library preparation steps. - 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.
7/2/2025DBC_9184_v114_revN_02Jul2025EnglishMinION
02 July 2025 - Updated overview of the protocol section within the introduction summary to a table format for library preparation steps. - 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.
7/2/2025DBC_9184_v114_revN_02Jul2025EnglishPromethION
02 July 2025 - Updated overview of the protocol section within the introduction summary to a table format for library preparation steps. - 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.
1/30/2025DBC_9184_v114_revM_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."
1/30/2025DBC_9184_v114_revM_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."
1/30/2025DBC_9184_v114_revM_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."
12/12/2024DBC_9184_v114_revL_12Dec2024EnglishGridION
12 December 2024 - Addition of link to MinION Mk1D IT requirements in "Compatibility of this 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".
12/12/2024DBC_9184_v114_revL_12Dec2024EnglishMinION
12 December 2024 - Addition of link to MinION Mk1D IT requirements in "Compatibility of this 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".
12/12/2024DBC_9184_v114_revL_12Dec2024EnglishPromethION
12 December 2024 - Addition of link to MinION Mk1D IT requirements in "Compatibility of this 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".
11/15/2024DBC_9184_v114_revK_20Nov2024EnglishGridION
November 2024 - Updated kit content information and image to include new kit format of SQK-NBD114.24 with plated native barcodes.
11/15/2024DBC_9184_v114_revK_20Nov2024EnglishMinION
November 2024 - Updated kit content information and image to include new kit format of SQK-NBD114.24 with plated native barcodes.
11/15/2024DBC_9184_v114_revK_20Nov2024EnglishPromethION
November 2024 - Updated kit content information and image to include new kit format of SQK-NBD114.24 with plated native barcodes.
8/21/2024DBC_9184_v114_revJ_22Mar2023EnglishGridION
21st August 2024 - Updated protocol introduction to new format. - Updated sample input recommendations to 100 ng for gDNA or amplicons. - Updated the 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.
8/21/2024DBC_9184_v114_revJ_22Mar2023EnglishMinION
21st August 2024 - Updated protocol introduction to new format. - Updated sample input recommendations to 100 ng for gDNA or amplicons. - Updated the 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.
8/21/2024DBC_9184_v114_revJ_22Mar2023EnglishPromethION
21st August 2024 - Updated protocol introduction to new format. - Updated sample input recommendations to 100 ng for gDNA or amplicons. - Updated the 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, 2024DBC_9184_v114_revI_22Mar2023EnglishPromethion Download 01 May 2024 - Updated snippet in 'Equipment and consumables' to no longer reference SQK-NBD114.96.
August 21st, 2024DBC_9184_v114_revI_22Mar2023EnglishGridion Download 01 May 2024 - Updated snippet in 'Equipment and consumables' to no longer reference SQK-NBD114.96.
August 21st, 2024DBC_9184_v114_revI_22Mar2023EnglishMinion Download 01 May 2024 - Updated snippet in 'Equipment and consumables' to no longer reference SQK-NBD114.96.
May 1st, 2024DBC_9184_v114_revH_22Mar2023EnglishMinion Download 07th March 2024 - Updated library loading recommendations to 35-50 fmols.
May 1st, 2024DBC_9184_v114_revH_22Mar2023EnglishPromethion Download 07th March 2024 - Updated library loading recommendations to 35-50 fmols.
May 1st, 2024DBC_9184_v114_revH_22Mar2023EnglishGridion Download 07th March 2024 - Updated library loading recommendations to 35-50 fmols.
March 7th, 2024DBC_9184_v114_revG_22Mar2023EnglishPromethion Download 21st 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. - Format change in "Native barcode ligation" step to fix downloaded checklist.
March 7th, 2024DBC_9184_v114_revG_22Mar2023EnglishGridion Download 21st 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. - Format change in "Native barcode ligation" step to fix downloaded checklist.
March 7th, 2024DBC_9184_v114_revG_22Mar2023EnglishMinion Download 21st 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. - Format change in "Native barcode ligation" step to fix downloaded checklist.
September 19th, 2023DBC_9184_v114_revE_22Mar2023EnglishPromethion 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, 2023DBC_9184_v114_revE_22Mar2023EnglishGridion 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, 2023DBC_9184_v114_revE_22Mar2023EnglishMinion 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, 2023DBC_9184_v114_revD_22Mar2023EnglishPromethion Download 1st August 2023 - Updated recommended magnetic rack
September 14th, 2023DBC_9184_v114_revD_22Mar2023EnglishGridion Download 1st August 2023 - Updated recommended magnetic rack
September 14th, 2023DBC_9184_v114_revD_22Mar2023EnglishMinion Download 1st August 2023 - Updated recommended magnetic rack
August 1st, 2023DBC_9184_v114_revC_22Mar2023EnglishPromethion Download May 2023 - Added amplicon tailed primer sequences to the introduction
August 1st, 2023DBC_9184_v114_revC_22Mar2023EnglishGridion Download May 2023 - Added amplicon tailed primer sequences to the introduction
August 1st, 2023DBC_9184_v114_revC_22Mar2023EnglishMinion Download May 2023 - Added amplicon tailed primer sequences to the introduction
April 28th, 2023DBC_9184_v114_revB_22Mar2023EnglishPromethion Download April 2023 - Added flow cell priming mix volumes to the "Priming and loading" steps
April 28th, 2023DBC_9184_v114_revB_22Mar2023EnglishGridion Download April 2023 - Added flow cell priming mix volumes to the "Priming and loading" steps
April 28th, 2023DBC_9184_v114_revB_22Mar2023EnglishMinion Download April 2023 - Added flow cell priming mix volumes to the "Priming and loading" steps
March 22nd, 2023DBC_9184_v114_revA_22Mar2023EnglishPromethion Download -
March 22nd, 2023DBC_9184_v114_revA_22Mar2023EnglishGridion Download -
March 22nd, 2023DBC_9184_v114_revA_22Mar2023EnglishMinion Download -

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