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[ccp4bb]: Cracking crystals and reams of advice
Title: Cracking crystals and reams of advice
Here’s a thought- grow your crystals in a small percentage of cryoprotectant, such as 2% glycerol (or 5-10% MPD). Then try to soak directly in high concentrations. Sometimes the protein binds the cryoprotectant molecule. Also, you should not have to soak for more than 5 minutes (less is OK- It’s only the outside that needs protection, not the inside) before flash cooling (Not freezing! Hugs and kisses to darling BR). It is a good idea to block the cryo-stream, or plunge into nitrogen and use cryo-tongs.
Or maybe your crystal is too large- a long bar might do with being cut. Choose the smallest loop possible, too. Check also to see if your poor diffraction is due to high mosaicity. Then follow the Oak Ridge Boys’ (More hugs and kisses all round there) method.
Lisa A. Nagy, Ph. D.
“Let me put my sunglasses on. That's the law around here, you got to wear your sunglasses. So you can feel cool.”