The following approaches can be helpful:
- To ensure good pore occupancy, 20ng of library DNA should be loaded for sequencing on MinION and GridION.
- Some carryover contaminants from extracted RNA samples or library prep reagents can negatively impact pore occupancy. To solve this issue, a 2x 1 ml 80% ethanol wash can be performed at step 1.6.19 instead of 1x 500 µl of 80% ethanol. Also, a cleaner library DNA can be prepared by performing an additional 250 µl SFB wash at step 1.8.8, for a total of 3x 250 µl SFB washes.