By Jack S. Cohen

The new paintings at the common incidence of antisense RNA and next use of antisense constructs to control gene expression has been of large price to molecular biologists and it presents the bulwark at the back of the endeavours mentioned during this ebook of the kingdom of the sphere.

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Of interest is that selective elimination of an ethyl group from deoxynucleoside in the mixed phosphite is enhanced by addition of excess lithium chloride. In the absence of lithium chloride, the major product is deoxythymidine 5'-diethylphosphoramidate. The alternative approach involves condensation of 5'-amino-5'-deoxy-3'-0-monomethoxytritylthymidine with 5'-0-trifluoroacetylamino-5'-deoxythymidine 3'-phosphate in the presence of triphenylphosphine-dipyridyldisulphide as activating agent (Greene and Letsinger, 1975).

The retention time increases with the number of negative charges on the oligonucleotide. In the case of oligonucleotides having the same length (size) substituted by various groups, the separation may be achieved by reverse-phase chromatography. , 1986b). ) (Asseline and Thuong, 1988a). P. P. P. P. P. Zbpd-(Nup)nbZ + (n- (1) Zbd-pNu --~2HObZ + (n - + nd-Nu (2) 1)d-Nu + HObZ (3) This method is used to prove the full deblocking of oligonucleotides (excluding any modifications which may occur at the nucleic base level Covalent Linking to Intercalating and Reactive Substances 37 during the oligonucleotide synthesis step) through a comparison of hydrolysates with the nucleoside unit samples or with previously synthesized modified nucleosides such as in the case of 5'-modified nucleosides dNu* (Equation 4), 3' -substitution via linkages other than the phosphodiester (Equation 5) or dinucleoside samples (in the case of compounds with modified internucleotide linkages) (Equation 6).

1988). Oxidation was completed using the appropriate amine in carbon tetrachloride as a onestep procedure following synthesis of oligodeoxynucleotides as Hphosphonates. , 1988). Using this procedure, the Hphosphonate and phosphite triester links are converted to the phosphoramidate and the phosphate triesters, respectively, as part of each synthesis cycle. This strategy therefore introduces the phosphate linkage as a neutral phosphate triester which is subsequently converted to the phosphate diester during deprotection of the final product.

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