RNA - synthesis, purification and crystallization.
Protocols for the routine chemical synthesis and purification of milligram quantities of RNA and DNA-RNA chimeras meeting the demands of X-ray crystallography are described. An efficient screening protocol to test the crystallizability of the molecules and the optimization of the crystallization conditions are presented, so as to allow reproduction by others. Essentially the same crystallization conditions as for DNA oligomers can be employed for RNA crystallization. Specific examples involving alternating octamers, G/C-rich decamers, sequences with overhangs, and drug complexes of chimeras are discussed. Success of the methods is attested by the crystals obtained which diffract to high resolution.
The Ohio State University, Laboratory of Biological Macromolecular Structure, Department of Biochemistry, Columbus 43210-1002, USA.
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MC
Wahl
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B
Ramakrishnan
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C
Ban
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X
Chen
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M
Sundaralingam