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SARS-CoV-2 virus PCR tiling and sequencing from RNA using EXP-MRT001 and SQK-RBK114.96 (MRT_9186_v114_revK_29Oct2025)

DateVersionLanguageDeviceChanges
10/29/2025MRT_9186_v114_revK_29Oct2025EnglishGridION
29th October 2025 - Section 3, 'Computer requirements and software' was removed and links for device IT requirements were added to section 1, '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. The subsection 'Check your flow cell' was moved from section 3 'Computer requirements and software' to section 2. - In section 2, 'Equipment and consumables' text in the SQK-RBK114.96 kit image was edited. - In sections 3, 4, and 5, text in images of plates were edited. - In section 5, 'Addition of rapid barcodes', a note was added to check your flow cell and to use the barcode wells only once. - In section 7, 'Priming and loading the MinION/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. A note was added to the end of this section to explain that when a flow cell is inserted into the MinION Mk1D, there will be a small gap around the sides of the device. - In 'Troubleshooting', sections on kit 9 chemistry and Guppy were removed as they are no longer supported.
10/29/2025MRT_9186_v114_revK_29Oct2025EnglishMinION
29th October 2025 - Section 3, 'Computer requirements and software' was removed and links for device IT requirements were added to section 1, '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. The subsection 'Check your flow cell' was moved from section 3 'Computer requirements and software' to section 2. - In section 2, 'Equipment and consumables' text in the SQK-RBK114.96 kit image was edited. - In sections 3, 4, and 5, text in images of plates were edited. - In section 5, 'Addition of rapid barcodes', a note was added to check your flow cell and to use the barcode wells only once. - In section 7, 'Priming and loading the MinION/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. A note was added to the end of this section to explain that when a flow cell is inserted into the MinION Mk1D, there will be a small gap around the sides of the device. - In 'Troubleshooting', sections on kit 9 chemistry and Guppy were removed as they are no longer supported.
6/23/2025MRT_9186_v114_revJ_23Jun2025EnglishGridION
23 June 2025 - Updated Data acquisition and basecalling section of the protocol to be compatible with updated versions of MinKNOW.
6/23/2025MRT_9186_v114_revJ_23Jun2025EnglishMinION
23 June 2025 - Updated Data acquisition and basecalling section of the protocol to be compatible with updated versions of MinKNOW.
12/13/2024MRT_9186_v114_revH_13Dec2024EnglishGridION
13 December 2024 - Addition of link to MinION Mk1D IT requirements in "Computer requirements and software" and updated link for EPI2ME. - 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. - Addition of MinION Mk1D user manual in "Data acquisition and basecalling", "How to start sequencing". - 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".
12/13/2024MRT_9186_v114_revH_13Dec2024EnglishMinION
13 December 2024 - Addition of link to MinION Mk1D IT requirements in "Computer requirements and software" and updated link for EPI2ME. - 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. - Addition of MinION Mk1D user manual in "Data acquisition and basecalling", "How to start sequencing". - 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".
9/17/2024MRT_9186_v114_revG_17Aug24EnglishGridION
17th Sept 2024 - Fixed table formatting in PCR Section of protcocol, step 1.
9/17/2024MRT_9186_v114_revG_17Aug24EnglishMinION
17th Sept 2024 - Fixed table formatting in PCR Section of protcocol, step 1.
9/17/2024MRT_9186_v114_revG_17Aug24EnglishGridION
17th August 2024 - Fixed table formatting in PCR Section of protcocol, step 1.
9/17/2024MRT_9186_v114_revG_17Aug24EnglishMinION
17th August 2024 - Fixed table formatting in PCR Section of protcocol, step 1.
9/17/2024MRT_9186_v114_revG_19Apr2023EnglishGridION
02nd July 2024 - Fixed table formatting in PCR Section of protcocol, step 1.
9/17/2024MRT_9186_v114_revG_19Apr2023EnglishMinION
02nd July 2024 - Fixed table formatting in PCR Section of protcocol, step 1.
7/2/2024MRT_9186_v114_revF_19Apr2023EnglishGridION
02nd July 2024 - Included the following statement to the introduction to the protocol and the Midnight primer sequence information: 'As mutations in SARS-CoV-2 variants emerge amplicon drop out may be observed; for users wishing to design their own primer spike-ins to address this we suggest adding to the appropriate primer pool at a final concentration between 3.33 µM and 6.66 µM.'
7/2/2024MRT_9186_v114_revF_19Apr2023EnglishMinION
02nd July 2024 - Included the following statement to the introduction to the protocol and the Midnight primer sequence information: 'As mutations in SARS-CoV-2 variants emerge amplicon drop out may be observed; for users wishing to design their own primer spike-ins to address this we suggest adding to the appropriate primer pool at a final concentration between 3.33 µM and 6.66 µM.'
July 2nd, 2024MRT_9186_v114_revE_19Apr2023EnglishGridion Download 24th January 2024 - Updated recommendations to take 11 µl of eluted DNA library into adapter addition and flow cell loading.
July 2nd, 2024MRT_9186_v114_revE_19Apr2023EnglishMinion Download 24th January 2024 - Updated recommendations to take 11 µl of eluted DNA library into adapter addition and flow cell loading.
January 24th, 2024MRT_9186_v114_revD_19Apr2023EnglishGridion Download 19th September 2023 - Updated flow cell loading instructions to include light shield installation
January 24th, 2024MRT_9186_v114_revD_19Apr2023EnglishMinion Download 19th September 2023 - Updated flow cell loading instructions to include light shield installation
September 19th, 2023MRT_9186_v114_revC_19Apr2023EnglishGridion Download 29th June 2023 - Reformatted the required MinKNOW settings in the "Data acquisition and basecalling" section.
September 19th, 2023MRT_9186_v114_revC_19Apr2023EnglishMinion Download 29th June 2023 - Reformatted the required MinKNOW settings in the "Data acquisition and basecalling" section.
May 3rd, 2023MRT_9186_v114_revB_19Apr2023EnglishGridion Download 3rd May - Updated the Midnight Primer sequences to include the V3 primer spike-ins
May 3rd, 2023MRT_9186_v114_revB_19Apr2023EnglishMinion Download 3rd May - Updated the Midnight Primer sequences to include the V3 primer spike-ins
April 19th, 2023MRT_9186_v114_revA_19Apr2023EnglishGridion Download -
April 19th, 2023MRT_9186_v114_revA_19Apr2023EnglishMinion Download -

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