Tonight I continued watching the Oxford Nanopore Learning Course on Metagenomics. I focused on reviewing the flow cell priming videos to learn some tricks. Hazel Johnson and Divya Mirrington demonstrated how to prime a flow cell. They highlighted the air pocket that needs to be removed by opening the priming port and removing a small amount of buffer. Then, 800 microliters priming solution are used to flush the flow cell. They recommend not adding the full amount to avoid bubbles. After five minutes, 200 microliters are added with the SpotON port open. Then, the library is added dropwise immediately. The priming and SpotON ports are closed to begin the run. The next video was a demonstration of priming the Flongle. First, the Flongle flow cell is inserted into the adapter. The cover is removed and pulled toward the lid. You can stick the tape on the lid during loading. Without air at the bottom of the pipette, the flush solution is added. The library is then added. The cover is placed to avoid evaporation, and the run is started. I learned by watching this video, as I have struggled with Flongles! It seems that it is ok to make contact with the port using the pipette tip. For the Flongle, you don’t remove buffer (and air pocket). I watched the Flongle video several times to remember how priming and loading is done. I hope this helps yield better results with the Flongle flow cells.
