Hi all,
My protein complex (120 kDa) has no symmetry. My manual picking is heavily biased as I can only recognise the complex under 1 single orientation (other orientations could be junk). Does anyone have advice on strategies for particle picking and template picking under these conditions?
Many thanks
Start with blob picking - that should allow you to identify other orientations from 2D classes, if they are present, which can then be used for template picking (or to train a model for Topaz/CrYOLO etc).
I sometimes use pre-trained model in Topaz Extract to start with, which seems to do a good job in many cases. This is my “un-biased” way of picking initial particles, especially when particle diameter is uncertain/unknown. After that, I remove junk with 2D classifications and use curated particles to do template-based picking or deep particle picking. Make sure to inspect particles picked by pre-trained Topaz Extract because on some micrographs/datasets, they do poor job.
As recommended above, blob picking is a great way to start as well.