Continuing with flow cell handling, the Human genome sequencing and analysis course from Nanopore Learning has a video entitled “A practical demonstration of priming and loading a flow cell.” In this short video, it is clear how the air pocket is removed with a P1000 set to 200 ul and slowly turned up to ~230. Removing too much buffer exposes the flow cell to air, damaging the pores. The flow cell is then flushed with 800 ul of buffer. After five minutes, 200 ul of a second flush are added. Immediately, the sample is added dropwise to the SpotON port. The priming and SpotON port are then closed. With new updates, ONT suggests adding the light shield before running.
Marta Verdugo from the Technical Services Team at Oxford Nanopore Technologies then presented a demo of setting up a new experiment with MinKNOW. They started a new experiment in MinKNOW and explained how the template can be used to run the same settings for an experiment that you perform routinely. I will keep that in mind. Kit selection is facilitated with the dynamic kit selection video. The default run options are 72 hours with bias voltage at 180 mV. Verdugo noted that the vias voltage can be adjusted for reusing flow cells. Basecalling models can be selected. Alignment to a reference can be performed in real-time. I haven’t tried this, though it may be useful for transcripts to a genome, for example. System Messages can be used to monitor available pores, warnings, and other messages. I am learning more about the options available in the software.
