January 2023
- Updated kit Flongle Flow Cell Priming kit name to Flongle Sequencing Expansion (EXP-FSE001) across protocol
- Changed the PromethION "Priming and loading the flow cell" section to the updated version.
January 2023
- Updated kit Flongle Flow Cell Priming kit name to Flongle Sequencing Expansion (EXP-FSE001) across protocol
- Changed the PromethION "Priming and loading the flow cell" section to the updated version.
January 2023
- Updated kit Flongle Flow Cell Priming kit name to Flongle Sequencing Expansion (EXP-FSE001) across protocol
- Changed the PromethION "Priming and loading the flow cell" section to the updated version.
January 2023
- Updated kit Flongle Flow Cell Priming kit name to Flongle Sequencing Expansion (EXP-FSE001) across protocol
- Changed the PromethION "Priming and loading the flow cell" section to the updated version.
18th October 2022
- Updated equipment and consumables to remove reference to the NEBNext Companion Module for Oxford Nanopore Technologies Ligation Sequencing
18th October 2022
- Updated equipment and consumables to remove reference to the NEBNext Companion Module for Oxford Nanopore Technologies Ligation Sequencing
18th October 2022
- Updated equipment and consumables to remove reference to the NEBNext Companion Module for Oxford Nanopore Technologies Ligation Sequencing
18th October 2022
- Updated equipment and consumables to remove reference to the NEBNext Companion Module for Oxford Nanopore Technologies Ligation Sequencing
22nd December 2021
In the Flongle branch of the protocol, "Equipment and consumables", added a note to the Flongle Sequencing Expansion information: "Oxford Nanopore Technologies deem the useful life of the Flow Cell Expansion to be 6 months from receipt by the customer."
22nd December 2021
In the Flongle branch of the protocol, "Equipment and consumables", added a note to the Flongle Sequencing Expansion information: "Oxford Nanopore Technologies deem the useful life of the Flow Cell Expansion to be 6 months from receipt by the customer."
22nd December 2021
In the Flongle branch of the protocol, "Equipment and consumables", added a note to the Flongle Sequencing Expansion information: "Oxford Nanopore Technologies deem the useful life of the Flow Cell Expansion to be 6 months from receipt by the customer."
22nd December 2021
In the Flongle branch of the protocol, "Equipment and consumables", added a note to the Flongle Sequencing Expansion information: "Oxford Nanopore Technologies deem the useful life of the Flow Cell Expansion to be 6 months from receipt by the customer."
22nd December 2021
In the Flongle branch of the protocol, "Equipment and consumables", added a note to the Flongle Sequencing Expansion information: "Oxford Nanopore Technologies deem the useful life of the Flow Cell Expansion to be 6 months from receipt by the customer."
22nd December 2021
In the Flongle branch of the protocol, "Equipment and consumables", added a note to the Flongle Sequencing Expansion information: "Oxford Nanopore Technologies deem the useful life of the Flow Cell Expansion to be 6 months from receipt by the customer."
22nd December 2021
In the Flongle branch of the protocol, "Equipment and consumables", added a note to the Flongle Sequencing Expansion information: "Oxford Nanopore Technologies deem the useful life of the Flow Cell Expansion to be 6 months from receipt by the customer."
22nd December 2021
In the Flongle branch of the protocol, "Equipment and consumables", added a note to the Flongle Sequencing Expansion information: "Oxford Nanopore Technologies deem the useful life of the Flow Cell Expansion to be 6 months from receipt by the customer."
7th October 2021
In the Flongle branch of the protocol, "Loading the Flongle flow cell":
- A clarifying image has been added under "The adapter needs to be plugged into your device, and the device should be plugged in and powered on before inserting the Flongle flow cell."
7th October 2021
In the Flongle branch of the protocol, "Loading the Flongle flow cell":
- A clarifying image has been added under "The adapter needs to be plugged into your device, and the device should be plugged in and powered on before inserting the Flongle flow cell."
7th October 2021
In the Flongle branch of the protocol, "Loading the Flongle flow cell":
- A clarifying image has been added under "The adapter needs to be plugged into your device, and the device should be plugged in and powered on before inserting the Flongle flow cell."
7th October 2021
In the Flongle branch of the protocol, "Loading the Flongle flow cell":
- A clarifying image has been added under "The adapter needs to be plugged into your device, and the device should be plugged in and powered on before inserting the Flongle flow cell."
23rd September 2021
In the Flongle branch of the protocol, "Loading the Flongle flow cell":
- A note has been added: "The adapter needs to be plugged into your device, and the device should be plugged in and powered on before inserting the Flongle flow cell."
- Flow cell loading instructions have been changed such that a 200 μl pipette is used to load the sequencing library into the flow cell.
23rd September 2021
In the Flongle branch of the protocol, "Loading the Flongle flow cell":
- A note has been added: "The adapter needs to be plugged into your device, and the device should be plugged in and powered on before inserting the Flongle flow cell."
- Flow cell loading instructions have been changed such that a 200 μl pipette is used to load the sequencing library into the flow cell.
23rd September 2021
In the Flongle branch of the protocol, "Loading the Flongle flow cell":
- A note has been added: "The adapter needs to be plugged into your device, and the device should be plugged in and powered on before inserting the Flongle flow cell."
- Flow cell loading instructions have been changed such that a 200 μl pipette is used to load the sequencing library into the flow cell.
23rd September 2021
In the Flongle branch of the protocol, "Loading the Flongle flow cell":
- A note has been added: "The adapter needs to be plugged into your device, and the device should be plugged in and powered on before inserting the Flongle flow cell."
- Flow cell loading instructions have been changed such that a 200 μl pipette is used to load the sequencing library into the flow cell.
22nd July 2021
In the Flongle branch of the protocol, "Adapter ligation and clean-up" step, a duplicated "important" note about flow cell loading recommendations has been removed.
22nd July 2021
In the Flongle branch of the protocol, "Adapter ligation and clean-up" step, a duplicated "important" note about flow cell loading recommendations has been removed.
22nd July 2021
In the Flongle branch of the protocol, "Adapter ligation and clean-up" step, a duplicated "important" note about flow cell loading recommendations has been removed.
22nd July 2021
In the Flongle branch of the protocol, "Adapter ligation and clean-up" step, a duplicated "important" note about flow cell loading recommendations has been removed.
24th June 2021
In the Flongle branch of the protocol, "Loading the Flongle flow cell" step, an amendment has been made to step 8: "Loading Beads II (LBII) mixed immediately before use, or Loading Solution (LS), if using. LS can be used instead of LBII when preparing libraries with the Ligation Sequencing Kit (SQK-LSK110)"
24th June 2021
In the Flongle branch of the protocol, "Loading the Flongle flow cell" step, an amendment has been made to step 8: "Loading Beads II (LBII) mixed immediately before use, or Loading Solution (LS), if using. LS can be used instead of LBII when preparing libraries with the Ligation Sequencing Kit (SQK-LSK110)"
24th June 2021
In the Flongle branch of the protocol, "Loading the Flongle flow cell" step, an amendment has been made to step 8: "Loading Beads II (LBII) mixed immediately before use, or Loading Solution (LS), if using. LS can be used instead of LBII when preparing libraries with the Ligation Sequencing Kit (SQK-LSK110)"
24th June 2021
In the Flongle branch of the protocol, "Loading the Flongle flow cell" step, an amendment has been made to step 8: "Loading Beads II (LBII) mixed immediately before use, or Loading Solution (LS), if using. LS can be used instead of LBII when preparing libraries with the Ligation Sequencing Kit (SQK-LSK110)"
10th June 2021
In the PromethION branch of the protocol, "Priming and loading the flow cell" step:
- Added a note to wait 20 minutes after taking the flow cells out of the fridge, and wiping off any condensation before inserting the flow cells into the device
- Added a note about correct insertion of the flow cell to avoid damaging the pins
10th June 2021
In the PromethION branch of the protocol, "Priming and loading the flow cell" step:
- Added a note to wait 20 minutes after taking the flow cells out of the fridge, and wiping off any condensation before inserting the flow cells into the device
- Added a note about correct insertion of the flow cell to avoid damaging the pins
10th June 2021
In the PromethION branch of the protocol, "Priming and loading the flow cell" step:
- Added a note to wait 20 minutes after taking the flow cells out of the fridge, and wiping off any condensation before inserting the flow cells into the device
- Added a note about correct insertion of the flow cell to avoid damaging the pins
10th June 2021
In the PromethION branch of the protocol, "Priming and loading the flow cell" step:
- Added a note to wait 20 minutes after taking the flow cells out of the fridge, and wiping off any condensation before inserting the flow cells into the device
- Added a note about correct insertion of the flow cell to avoid damaging the pins
21st April 2021
In the Flongle branch of the protocol:
- "Equipment and consumables", "DNA repair and end-prep" and "Loading the Flongle flow cell" steps: added additional information about flow cell deterioration/saturation, and updated the Flongle Sequencing Expansion information to mention Loading Solution (LS) and reduction of flow cell deterioration when storing components in glass vials instead of plastic
- "Adapter ligation and clean-up" step: updated the loading recommendations to "Following standard input recommendations, the protocol should produce enough final library (adapted DNA in EB) to load at least two Flongle flow cells. We recommend reserving enough library to load a second Flongle flow cell. We recommend loading 3-20 fmol of this final prepared library onto the flow cell. Loading more than 50 fmol can have a detrimental effect on throughput. Dilute the library in EB or nuclease free water up to a final volume of 5 μl."
- "Loading the Flongle flow cell" step: added the warning "ALWAYS wear gloves when handling Flongle flow cells and adapters to avoid damage to the flow cell or adapter."
Inclusion of the optional Adapter Mix II Expansion (EXP-AMII001) kit for instances when a user is running a subset of barcodes in one experiment and needs extra Adapter Mix II for more reactions:
- EXP-AMII001 is included in the "Equipment and consumables" section
- Explanation of when to use EXP-AMII001 in the "Adapter ligation and clean-up" step
21st April 2021
In the Flongle branch of the protocol:
- "Equipment and consumables", "DNA repair and end-prep" and "Loading the Flongle flow cell" steps: added additional information about flow cell deterioration/saturation, and updated the Flongle Sequencing Expansion information to mention Loading Solution (LS) and reduction of flow cell deterioration when storing components in glass vials instead of plastic
- "Adapter ligation and clean-up" step: updated the loading recommendations to "Following standard input recommendations, the protocol should produce enough final library (adapted DNA in EB) to load at least two Flongle flow cells. We recommend reserving enough library to load a second Flongle flow cell. We recommend loading 3-20 fmol of this final prepared library onto the flow cell. Loading more than 50 fmol can have a detrimental effect on throughput. Dilute the library in EB or nuclease free water up to a final volume of 5 μl."
- "Loading the Flongle flow cell" step: added the warning "ALWAYS wear gloves when handling Flongle flow cells and adapters to avoid damage to the flow cell or adapter."
Inclusion of the optional Adapter Mix II Expansion (EXP-AMII001) kit for instances when a user is running a subset of barcodes in one experiment and needs extra Adapter Mix II for more reactions:
- EXP-AMII001 is included in the "Equipment and consumables" section
- Explanation of when to use EXP-AMII001 in the "Adapter ligation and clean-up" step
21st April 2021
In the Flongle branch of the protocol:
- "Equipment and consumables", "DNA repair and end-prep" and "Loading the Flongle flow cell" steps: added additional information about flow cell deterioration/saturation, and updated the Flongle Sequencing Expansion information to mention Loading Solution (LS) and reduction of flow cell deterioration when storing components in glass vials instead of plastic
- "Adapter ligation and clean-up" step: updated the loading recommendations to "Following standard input recommendations, the protocol should produce enough final library (adapted DNA in EB) to load at least two Flongle flow cells. We recommend reserving enough library to load a second Flongle flow cell. We recommend loading 3-20 fmol of this final prepared library onto the flow cell. Loading more than 50 fmol can have a detrimental effect on throughput. Dilute the library in EB or nuclease free water up to a final volume of 5 μl."
- "Loading the Flongle flow cell" step: added the warning "ALWAYS wear gloves when handling Flongle flow cells and adapters to avoid damage to the flow cell or adapter."
Inclusion of the optional Adapter Mix II Expansion (EXP-AMII001) kit for instances when a user is running a subset of barcodes in one experiment and needs extra Adapter Mix II for more reactions:
- EXP-AMII001 is included in the "Equipment and consumables" section
- Explanation of when to use EXP-AMII001 in the "Adapter ligation and clean-up" step
21st April 2021
In the Flongle branch of the protocol:
- "Equipment and consumables", "DNA repair and end-prep" and "Loading the Flongle flow cell" steps: added additional information about flow cell deterioration/saturation, and updated the Flongle Sequencing Expansion information to mention Loading Solution (LS) and reduction of flow cell deterioration when storing components in glass vials instead of plastic
- "Adapter ligation and clean-up" step: updated the loading recommendations to "Following standard input recommendations, the protocol should produce enough final library (adapted DNA in EB) to load at least two Flongle flow cells. We recommend reserving enough library to load a second Flongle flow cell. We recommend loading 3-20 fmol of this final prepared library onto the flow cell. Loading more than 50 fmol can have a detrimental effect on throughput. Dilute the library in EB or nuclease free water up to a final volume of 5 μl."
- "Loading the Flongle flow cell" step: added the warning "ALWAYS wear gloves when handling Flongle flow cells and adapters to avoid damage to the flow cell or adapter."
Inclusion of the optional Adapter Mix II Expansion (EXP-AMII001) kit for instances when a user is running a subset of barcodes in one experiment and needs extra Adapter Mix II for more reactions:
- EXP-AMII001 is included in the "Equipment and consumables" section
- Explanation of when to use EXP-AMII001 in the "Adapter ligation and clean-up" step
2nd February 2021
In the Flongle branch of the protocol:
- "Equipment and consumables" step: updated Flongle Sequencing Expansion (EXP-FSE001) kit contents
- "Loading the Flongle flow cell" step: updated instructions. SBII, LBII and FB should be used from the Flongle Sequencing Expansion instead of SQB, LB and FB from the sequencing kit
2nd February 2021
In the Flongle branch of the protocol:
- "Equipment and consumables" step: updated Flongle Sequencing Expansion (EXP-FSE001) kit contents
- "Loading the Flongle flow cell" step: updated instructions. SBII, LBII and FB should be used from the Flongle Sequencing Expansion instead of SQB, LB and FB from the sequencing kit
2nd February 2021
In the Flongle branch of the protocol:
- "Equipment and consumables" step: updated Flongle Sequencing Expansion (EXP-FSE001) kit contents
- "Loading the Flongle flow cell" step: updated instructions. SBII, LBII and FB should be used from the Flongle Sequencing Expansion instead of SQB, LB and FB from the sequencing kit
2nd February 2021
In the Flongle branch of the protocol:
- "Equipment and consumables" step: updated Flongle Sequencing Expansion (EXP-FSE001) kit contents
- "Loading the Flongle flow cell" step: updated instructions. SBII, LBII and FB should be used from the Flongle Sequencing Expansion instead of SQB, LB and FB from the sequencing kit
2nd October 2020
Updated Flongle equipment and flow cell loading instructions in line with the release of the Flongle Flow Cell Priming Kit (EXP-FLP003)
2nd October 2020
Updated Flongle equipment and flow cell loading instructions in line with the release of the Flongle Flow Cell Priming Kit (EXP-FLP003)
2nd October 2020
Updated Flongle equipment and flow cell loading instructions in line with the release of the Flongle Flow Cell Priming Kit (EXP-FLP003)
2nd October 2020
Updated Flongle equipment and flow cell loading instructions in line with the release of the Flongle Flow Cell Priming Kit (EXP-FLP003)
28th January 2020
Added an optional instruction in the PromethION branch of the protocol for setting aside 1 µl of library for performing a QC experiment using Flongle
28th January 2020
Added an optional instruction in the PromethION branch of the protocol for setting aside 1 µl of library for performing a QC experiment using Flongle
28th January 2020
Added an optional instruction in the PromethION branch of the protocol for setting aside 1 µl of library for performing a QC experiment using Flongle
28th January 2020
Added an optional instruction in the PromethION branch of the protocol for setting aside 1 µl of library for performing a QC experiment using Flongle
23rd January 2020
Replaced DNA input recommendations for R10 flow cells with R10.3:
- starting with 1.5-3 µg (or 150-300 fmol) DNA
- loading 25-75 fmols onto the flow cell
23rd January 2020
Replaced DNA input recommendations for R10 flow cells with R10.3:
- starting with 1.5-3 µg (or 150-300 fmol) DNA
- loading 25-75 fmols onto the flow cell
23rd January 2020
Replaced DNA input recommendations for R10 flow cells with R10.3:
- starting with 1.5-3 µg (or 150-300 fmol) DNA
- loading 25-75 fmols onto the flow cell
23rd January 2020
Replaced DNA input recommendations for R10 flow cells with R10.3:
- starting with 1.5-3 µg (or 150-300 fmol) DNA
- loading 25-75 fmols onto the flow cell
30th September 2019
The optional DNA fragmentation step has been removed from the protocol, since it is now available as a [stand-alone protocol](https://community.nanoporetech.com/extraction_methods/dna-fragmentation).
30th September 2019
The optional DNA fragmentation step has been removed from the protocol, since it is now available as a [stand-alone protocol](https://community.nanoporetech.com/extraction_methods/dna-fragmentation).
30th September 2019
The optional DNA fragmentation step has been removed from the protocol, since it is now available as a [stand-alone protocol](https://community.nanoporetech.com/extraction_methods/dna-fragmentation).
30th September 2019
The optional DNA fragmentation step has been removed from the protocol, since it is now available as a [stand-alone protocol](https://community.nanoporetech.com/extraction_methods/dna-fragmentation).
20th September 2019
- A note has been added at the end of the library preparation instructions about using the Sequencing Auxiliary Vials (EXP-AUX001) for splitting a library into two aliquots
20th September 2019
- A note has been added at the end of the library preparation instructions about using the Sequencing Auxiliary Vials (EXP-AUX001) for splitting a library into two aliquots
20th September 2019
- A note has been added at the end of the library preparation instructions about using the Sequencing Auxiliary Vials (EXP-AUX001) for splitting a library into two aliquots
20th September 2019
- A note has been added at the end of the library preparation instructions about using the Sequencing Auxiliary Vials (EXP-AUX001) for splitting a library into two aliquots
14th August 2019
The protocol has been expanded to include instructions for library prep on multiple nanopore sequencing devices. The user can select instructions specific to their device by selecting from the dropdown menu on the right hand side of the protocol.
14th August 2019
The protocol has been expanded to include instructions for library prep on multiple nanopore sequencing devices. The user can select instructions specific to their device by selecting from the dropdown menu on the right hand side of the protocol.
14th August 2019
The protocol has been expanded to include instructions for library prep on multiple nanopore sequencing devices. The user can select instructions specific to their device by selecting from the dropdown menu on the right hand side of the protocol.
14th August 2019
The protocol has been expanded to include instructions for library prep on multiple nanopore sequencing devices. The user can select instructions specific to their device by selecting from the dropdown menu on the right hand side of the protocol.
19th July 2019
Protocol structure has been condensed to remove sections about DNA/RNA QC, software installation and use, and flow cell check. These sections are available as stand-alone documents, which are linked out from the protocol. The checklist protocol is now available for download from the button in the protocol header
Updated with new information for MinKNOW release v19.06:
- Compression of queued reads
- Pause functionality
- UI improvements including:
- Flow cell health at platform QC
- Flow cell status on landing page
- R10 flow cell running scripts
- New Fast Models for RNA and DNA
- PDF report and CSV files for Duty time and Cumulative output are now found in same location as actual reads and fastQ files
- Flongle hardware check for confirming Flongle adapter functions correctly
Update to the flow cell priming section, and instructions for topping up the flow cell with fuel, following the release of the Flow Cell Priming Kit (EXP-FLP002)
Updates to kit contents with the introduction of the Control Expansion kit (EXP-CTL001) and removal of the Lambda Control DNA from the Ligation Sequencing Kit