Crystallization of the hairpin ribozyme: illustrative protocols.
Conditions and techniques that result in successful crystallization differ from RNA to RNA. However, there are some general principles that facilitate crystallization of most RNAs. Three procedures that were instrumental in obtaining well-ordered crystals of the hairpin ribozyme are described in this chapter. These are: i) the design of a series of candidate crystallization constructs; ii) the evaluation of conditions to obtain monodisperse RNA; and iii) the use of seeding techniques to separate nucleation and growth events during crystallization. These procedures can be usefully adapted for the crystallization of other RNAs.
Rupert
Peter B
PB
Division of Basic Sciences, Fred Hutchinson Cancer Research Center, Seattle, WA, USA.
Ferré-D'Amaré
Adrian R
AR
D001665:Binding Sites; D003460:Crystallization; D018360:Crystallography, X-Ray / Q000379:methods; D009690:Nucleic Acid Conformation; D016133:Polymerase Chain Reaction / Q000379:methods; D016337:RNA, Catalytic / Q000096:biosynthesis / Q000737:chemistry*; D019914:Taq Polymerase
D016337:RNA, Catalytic; C418481:hairpin ribozyme; D019914:Taq Polymerase