A method to produce microseed stock for use in the crystallization of biological macromolecules.
A method is presented for producing a seed-stock mixture for macromolecular crystallization. A PTFE bead and micro-centrifuge tube act as mortar and pestle for pulverizing seed crystals of macromolecules. Energy for the bead's motion is supplied by a vortex mixer or an ultrasonic bath. The crushed crystal is serially diluted to prepare a seed-stock mixture of the desired concentration for crystallization. Crystals produced using both hanging-drop vapor diffusion and a capillary microbatch method show expected dilution behavior. This technique of producing seed stock is compared with traditional means and advantages over the standard protocol are demonstrated.
Luft
J R
JR
Hauptman-Woodward Medical Research Institute, 73 High Street, Buffalo, New York 14203-1196, USA. luft@hwi.buffalo.edu
DeTitta
G T
GT
D001671:Biochemistry / Q000379:methods*; D003460:Crystallization; D004058:Diffusion; D005898:Glass; D046911:Macromolecular Substances; D008967:Molecular Biology / Q000379:methods; D011506:Proteins / Q000737:chemistry*; D014465:Ultrasonics
D046911:Macromolecular Substances; D011506:Proteins