From portable to high-throughput benchtop devices, real-time nanopore sensing is a new generation of technology uncovering new biology across multiple sectors.
14th 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 5, 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' and 'Flongle Flow Cell loading', 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 V14 (SQK-LSK114) on the Kit page'. And the section on 'Duplex basecalling' was removed as it is no longer applicable.
- For the Flongle branch, the sections on 'Flow cell deterioration/saturation' and 'Flongle Sequencing Expansion' were edited.
- The video link for priming and loading a Flongle Flow Cell was updated in section 5, step 4.
- Screenshots were removed in section 6, 'Data acquisition and basecalling'.
5/14/2025
CDE_9162_v114_revW_14May2025
English
GridION
14th 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 5, 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' and 'Flongle Flow Cell loading', 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 V14 (SQK-LSK114) on the Kit page'. And the section on 'Duplex basecalling' was removed as it is no longer applicable.
- For the Flongle branch, the sections on 'Flow cell deterioration/saturation' and 'Flongle Sequencing Expansion' were edited.
- The video link for priming and loading a Flongle Flow Cell was updated in section 5, step 4.
- Screenshots were removed in section 6, 'Data acquisition and basecalling'.
5/14/2025
CDE_9162_v114_revW_14May2025
English
MinION
14th 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 5, 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' and 'Flongle Flow Cell loading', 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 V14 (SQK-LSK114) on the Kit page'. And the section on 'Duplex basecalling' was removed as it is no longer applicable.
- For the Flongle branch, the sections on 'Flow cell deterioration/saturation' and 'Flongle Sequencing Expansion' were edited.
- The video link for priming and loading a Flongle Flow Cell was updated in section 5, step 4.
- Screenshots were removed in section 6, 'Data acquisition and basecalling'.
5/14/2025
CDE_9162_v114_revW_14May2025
English
PromethION
14th 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 5, 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' and 'Flongle Flow Cell loading', 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 V14 (SQK-LSK114) on the Kit page'. And the section on 'Duplex basecalling' was removed as it is no longer applicable.
- For the Flongle branch, the sections on 'Flow cell deterioration/saturation' and 'Flongle Sequencing Expansion' were edited.
- The video link for priming and loading a Flongle Flow Cell was updated in section 5, step 4.
- Screenshots were removed in section 6, 'Data acquisition and basecalling'.
5/14/2025
CDE_9162_v114_revW_14May2025
English
Flongle
14th 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 5, 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' and 'Flongle Flow Cell loading', 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 V14 (SQK-LSK114) on the Kit page'. And the section on 'Duplex basecalling' was removed as it is no longer applicable.
- For the Flongle branch, the sections on 'Flow cell deterioration/saturation' and 'Flongle Sequencing Expansion' were edited.
- The video link for priming and loading a Flongle Flow Cell was updated in section 5, step 4.
- Screenshots were removed in section 6, 'Data acquisition and basecalling'.
5/14/2025
CDE_9162_v114_revW_14May2025
English
GridION
14th 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 5, 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' and 'Flongle Flow Cell loading', 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 V14 (SQK-LSK114) on the Kit page'. And the section on 'Duplex basecalling' was removed as it is no longer applicable.
- For the Flongle branch, the sections on 'Flow cell deterioration/saturation' and 'Flongle Sequencing Expansion' were edited.
- The video link for priming and loading a Flongle Flow Cell was updated in section 5, step 4.
- Screenshots were removed in section 6, 'Data acquisition and basecalling'.
5/14/2025
CDE_9162_v114_revW_14May2025
English
MinION
14th 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 5, 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' and 'Flongle Flow Cell loading', 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 V14 (SQK-LSK114) on the Kit page'. And the section on 'Duplex basecalling' was removed as it is no longer applicable.
- For the Flongle branch, the sections on 'Flow cell deterioration/saturation' and 'Flongle Sequencing Expansion' were edited.
- The video link for priming and loading a Flongle Flow Cell was updated in section 5, step 4.
- Screenshots were removed in section 6, 'Data acquisition and basecalling'.
5/14/2025
CDE_9162_v114_revW_14May2025
English
PromethION
14th 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 5, 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' and 'Flongle Flow Cell loading', 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 V14 (SQK-LSK114) on the Kit page'. And the section on 'Duplex basecalling' was removed as it is no longer applicable.
- For the Flongle branch, the sections on 'Flow cell deterioration/saturation' and 'Flongle Sequencing Expansion' were edited.
- The video link for priming and loading a Flongle Flow Cell was updated in section 5, step 4.
- Screenshots were removed in section 6, 'Data acquisition and basecalling'.
1/30/2025
CDE_9162_v114_revV_30Jan2025
English
Flongle
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."
1/30/2025
CDE_9162_v114_revV_30Jan2025
English
GridION
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."
1/30/2025
CDE_9162_v114_revV_30Jan2025
English
MinION
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."
1/30/2025
CDE_9162_v114_revV_30Jan2025
English
PromethION
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."
12/12/2024
CDE_9162_v114_revU_12Dec2024
English
Flongle
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".
12/12/2024
CDE_9162_v114_revU_12Dec2024
English
GridION
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".
12/12/2024
CDE_9162_v114_revU_12Dec2024
English
MinION
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".
12/12/2024
CDE_9162_v114_revU_12Dec2024
English
PromethION
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".
12/12/2024
CDE_9162_v114_revU_12Dec2023
English
Flongle
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".
12/12/2024
CDE_9162_v114_revU_12Dec2023
English
GridION
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".
12/12/2024
CDE_9162_v114_revU_12Dec2023
English
MinION
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".
12/12/2024
CDE_9162_v114_revU_12Dec2023
English
PromethION
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".
8/21/2024
CDE_9162_v114_revT_29Jun2022
English
Flongle
21st August 2024
- 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.
8/21/2024
CDE_9162_v114_revT_29Jun2022
English
GridION
21st August 2024
- 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.
8/21/2024
CDE_9162_v114_revT_29Jun2022
English
MinION
21st August 2024
- 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.
8/21/2024
CDE_9162_v114_revT_29Jun2022
English
PromethION
21st August 2024
- 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.
08 Aug 2024
Added sentence to give LMD fragment length in Introduction to the protocol/Equipment and consumables/Control Expansion (EXP-CTL001) contents
"Lambda DNA (LMD) fragment length is 48.5 kb."
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
18th April 2024
- Updated elution step in "Adapter ligation and clean-up" section to specify 10 minute incubation for PromethION branch.
08 Aug 2024
Added sentence to give LMD fragment length in Introduction to the protocol/Equipment and consumables/Control Expansion (EXP-CTL001) contents
"Lambda DNA (LMD) fragment length is 48.5 kb."
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
18th April 2024
- Updated elution step in "Adapter ligation and clean-up" section to specify 10 minute incubation for PromethION branch.
08 Aug 2024
Added sentence to give LMD fragment length in Introduction to the protocol/Equipment and consumables/Control Expansion (EXP-CTL001) contents
"Lambda DNA (LMD) fragment length is 48.5 kb."
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
18th April 2024
- Updated elution step in "Adapter ligation and clean-up" section to specify 10 minute incubation for PromethION branch.
08 Aug 2024
Added sentence to give LMD fragment length in Introduction to the protocol/Equipment and consumables/Control Expansion (EXP-CTL001) contents
"Lambda DNA (LMD) fragment length is 48.5 kb."
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
18th April 2024
- Updated elution step in "Adapter ligation and clean-up" section to specify 10 minute incubation for PromethION branch.
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.
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.
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.
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.
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
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
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
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