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By John N. Abelson, Melvin I. Simon, David M.J. Lilley, James E. Dahlberg

To appreciate the buildings and dynamics of DNA and its interactions with ligands, an entire research of its confirmational flexibility is needed. This quantity and its spouse, quantity 212 of Methods in Enzymology, collate in a single resource all of the tools and methods had to do so aim

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4 g) is placed in a l-liter flask with 400 ml anhydrous ether. 3 Eq). 5 hr. Stirring is maintained for an additional 1 hr and then stopped in order to allow precipitates to settle. The reaction mixture is filtered through a medium frit sintered glass funnel, and the salts are washed with an additional 50 ml anhydrous ether. Ether and trimethylsilyl chloride are removed from the filtrate by rotary evaporation to give ap- 16 CHEMICALSYNTHESISOF DNA [ 1] proximately 100 ml of crude phosphine. 8]. Synthesis of Deoxynucleoside 3'-Phosphorothioamidites Protected Deoxynucleosides.

7. IRL Press at Oxford Univ. Press, Oxford, 1989. [2] PURIFICATION OF SYNTHETICDNA 29 J FIG. 2. Preparative anion-exchangeHPLC chromatogram of the crude self-complementary dodecamer d(CGCGAATTCGCG),one 1/tmol synthesis. The sharp peak is benzamide, followedby failure sequences,and the final major peak is the desired product. The chromatogram was monitored at 280 nm with a fixed wavelength ultraviolet detector. Trityl-off R e v e r s e d - P h a s e Chromatographic Purification Trityl-off reversed-phase H P L C is the final H P L C purification step and follows either anion-exchange or trityl-on reversed-phase HPLC.

L. Gatfney and R. A. Jones, Tetrahedron Lett. 23, 2257 (1982). 12B. L. Gaffney, L. A. Marky, and R. A. Jones, Tetrahedron 40, 3 (1984). 13L. W. McLaughlin, T. Leong, F. Benseler, and N. Piel, Nucleic Acids Res. 16, 5631 (1988). 2c,~o"~'~y X c,c,2c,2o" ~',~ = XI FiG. 2. Preparation of derivatives of d2~p and d6SG suitable for oligodeoxynucleotide synthesis. This figure should be used in conjunction with procedures l, 2, and 3. Reagents: 1, benzoic anhydride; 2, 2,4,6-triisopropylbenzcnesulfonyl chloride; 3, hydrazine; 4, Ag20; 5, NaOH (brief); 6, Li2S; 7, bromopropionitrile; 8, dimethoxytrityl chloride, then cyanoethylN,N-diisopropylchlorophosphoramidite.

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