Crystal growth: Tips and Tricks: Difference between revisions

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Crystal growth: Tips and Tricks (edit)

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→‎I tried Hampton Screen I & II with my protein and have no crystals. What can I do?
 
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#* [http://www.emeraldbiosystems.com Emerald Biosystems]
#* [http://www.emeraldbiosystems.com Emerald Biosystems]
#* [http://www.jenabioscience.com/cms/en/1/browse/632_screens.html Jena Bioscience]
#* [http://www.jenabioscience.com/cms/en/1/browse/632_screens.html Jena Bioscience]
#* [[Crystallization screens and methods | Other screens ]]
#* [http://www.moleculardimensions.com Molecular Dimensions]
#* See also [[Crystallization screens and methods]]
# [[Modifying_the_protein_to_crystallize_better | Modify your protein. ]]
# [[Modifying_the_protein_to_crystallize_better | Modify your protein. ]]


==Improving crystals==
==Improving crystals==
===Reducing the mosaicity===
===Reducing the mosaicity===
Things to try in the order crystallization-related / cryo-related / geometry-related:
 
# change precipitant, pH, temperature; try additives
Things to try concerning crystallization - see also [[Improving crystal quality]].
# try co-crystallization with glycerol (2%, 5% or 10%).
For cryo question see [[cryo]] and [[Data collection: Tips and Tricks]].
# incubate your crystal longer in your cryo/stabilization solution.
 
# optimize the cryo conditions
* change precipitant, pH, temperature; try additives
# anneal (freeze/thaw) the crystal - '''DISCLAIMER''': your mosaicity can go up as well as down with annealing
* try co-crystallization with glycerol (2%, 5% or 10%).
# the mosaicity may be anisotropic, so try to orient the crystal in the loop such that you can shoot along a different axis
* incubate your crystal longer in your cryo/stabilization solution.
# try smaller crystals (often growth faults lead to end of crystal growth)
* optimize the cryo conditions
# use a synchrotron beam (as it is less divergent than the usual home source, and the apparent mosaicity is the sum of the crystal mosaicity and the crossfire)
* anneal (freeze/thaw) the crystal - '''DISCLAIMER''': your mosaicity can go up as well as down with annealing
# shoot at parts of the crystal to find if there are good and bad parts (this requires a small beam)
* the mosaicity may be anisotropic, so try to orient the crystal in the loop such that you can shoot along a different axis
# look at Elspeth Garman's papers, and/or check her talks at RapiData workshops (anybody with a link??)
* look at Elspeth Garman's papers, and/or check her talks at RapiData workshops (anybody with a link??)


==References==
==References==
* J.W. Pflugrath: Macromolecular cryocrystallography—methods for cooling and mounting protein crystals at cryogenic temperatures http://dx.doi.org/10.1016/j.ymeth.2004.03.032
* J.W. Pflugrath (2004) Macromolecular cryocrystallography—methods for cooling and mounting protein crystals at cryogenic temperatures. ''Methods'' '''34''', 415-423 http://dx.doi.org/10.1016/j.ymeth.2004.03.032
* papers listed at http://www.px.nsls.bnl.gov/courses/papers/ZD_EG_papers.html
* H.D. Bellamy, E.H. Snell, J. Lovelace, M. Pokross and G.E.O. Borgstahl (2000) The high-mosaicity illusion: revealing the true physical characteristics of macromolecular crystals. ''Acta Cryst.'' '''D56''', 986–995
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