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This purification step is critically important. One must ensure that the desired ribonucleoside 2′-acetal is not contaminated with any 3′-acetal to prevent subsequent incorporation of 2′→5′, internucleotidic phosphodiester linkages into the oligoribonucleotide during solid-phase synthesis
-
This purification step is critically important. One must ensure that the desired ribonucleoside 2′-acetal is not contaminated with any 3′-acetal to prevent subsequent incorporation of (2′→5′)- internucleotidic phosphodiester linkages into the oligoribonucleotide during solid-phase synthesis.
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The RP-HPLC retention time of 29 (tR, 19.6 min) is identical to that of an authentic sample of 4-(N-methylamino)benzyl alcohol that was prepared from the reduction of 4-(N-methylamino)benzoic acid see ref 25
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R = 19.6 min) is identical to that of an authentic sample of 4-(N-methylamino)benzyl alcohol that was prepared from the reduction of 4-(N-methylamino)benzoic acid (see ref 25).
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Synthetic (3′→5′)UpU is a substrate for BSP but synthetic (2′→5′)UpU is not. Data are presented in the Supporting Information. See also: (a) Giannaris, P. A.; Damha, M. J. Nucleic Acids Res. 1993, 21, 4742-4749.
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Commercial bovine spleen phosphodiesterase contains some (1% w/w) adenosine deaminase (ADA) as a contaminant. Such a low concentration of ADA in the digestion reaction was sufficient to quantitatively convert adenosine to inosine. This conversion was unambiguously demonstrated by mixing adenosine with BSP/BAP under the conditions used for the digestion of the RNA oligomer. RP-HPLC analysis of the reaction indicated complete conversion of adenosine to inosine, which had a retention time identical to that of a commercial sample of inosine (data shown in the Supporting Information).
-
Commercial bovine spleen phosphodiesterase contains some (1% w/w) adenosine deaminase (ADA) as a contaminant. Such a low concentration of ADA in the digestion reaction was sufficient to quantitatively convert adenosine to inosine. This conversion was unambiguously demonstrated by mixing adenosine with BSP/BAP under the conditions used for the digestion of the RNA oligomer. RP-HPLC analysis of the reaction indicated complete conversion of adenosine to inosine, which had a retention time identical to that of a commercial sample of inosine (data shown in the Supporting Information).
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55
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2 is employed as an eluent.
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2 is employed as an eluent.
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