I am attempting to perform disk diffusion assays with novel extracts but the blank disks I use (remel, processed blank disks) are producing small zones of inhibition. I have tried using disks from whatman filter paper but those too leaves a zone. When using methanol as a negative control the disks seem dry but I get a zone larger than a blank disk but with high variability. I have tried drying the disk from 15 minutes to 2 hours in a flow hood but that seems to have little effect on the zone diameter. Could the issue be the bacteria I am using or some other issue?
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