Polymer support oligonucleotide synthesis XVIII1.2): use of β-cyanoethyi-N,N-dialkylamino-/N-morpholino phosphoramidite of deoxynucleosides for the synthesis of DNA fragments simplifying deprotection and isolation of the final product

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78

作者:

D N.SinhaJ.BiernatJ.McManusH.Köster

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摘要:

Various 5′-0-N-protected deoxynucleoside-3′-0-β-cyanoethyl-N,N-dialkylamino-/N-morpholinophosphoramidites were prepared from β-cyanoethyl monochlorophosphoramidites of N,N-dimethylamine, N,N-diisopropylamine and N-morpholine. These active deoxynucleoside phosphites have successfully been used for oligodeoxy-nucleotide synthessis on controlled pore glass as polymer support and are very suitable for automated DNA-synthesis due to their stability in solution. The intermediate dichloro-β-cyanoethoxy-phosphine can easily be prepared free from any PCl<sub>3</sub> contamination. The active monomers obtained from β-cyanoethyl monochloro N,N-diisopropylaminophosphoramidites are favoured. Cleavage of the oligonucleotide chain from the polymer support, N-deacylation and deprotection of β-cyanoethyl group from the phosphate triester moiety can be performed in one step with concentrated aqueous ammonia. Mixed oligodeoxynucleotides are characterized by the sequencing method of Maxam and Gilbert.

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DOI:

10.1093/nar/12.11.4539

被引量:

520

年份:

1984

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1997
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