17th November 2025
- Following the recent release of MinKNOW 25.09, this protocol was updated so that live basecalling can be performed with this latest version of MinKNOW.
- The sections on 'Data acquisition' and 'Basecalling and downstream analysis' were updated to reflect this change.
9/29/2025
CAS_9226_v114_revB_29Sep2025
English
PromethION
29th September 2025
- In 'Overview of the protocol', in the Table under 'Library preparation', the timing for priming and loading a flow cell was increased from 5 min to 10 min.
- In 'Equipment and consumables', the image of the LSK114 kit in single-use tube format was removed.
- In section 8, 'DNA repair and end-prep', the step to 'Check your flow cell', the link for the flow cell check in the MinKNOW protocol was replaced with the link for the flow cell check document. In step 1, point 4, the FFPE DNA Repair Buffer and Ultra II End-prep Reaction Buffer were removed as reagents from the Companion Module v2 are more efficient for library preparation compared with these reagent from the previous Companion module. In step 15, instructions were provided on how to resuspend the AMPure XP bead pellet with nuclease-free water.
- In section 10, 'Priming and loading the flow cell', the instruction for removing the flow cell out of the fridge was moved to the top of this section. Instructions for the single-use tube format of the LSK114 kit were removed from step 2 and a step to 'Complete a flow cell check' was included.
- In 'Troubleshooting', section 16, the video links for priming and loading a MinION Flow Cell were replaced with links for the PromethION Flow Cell.
