Polymer support oligonucleotide synthesis XVIII^1.2): use ofβ-cyanoethyi-N,N-dialkylamino-/N-morpholino phosphoramidite of deoxynucleosides for the synthesis of DNA fragments simplifying deprotection and isolation of the final product

Abstract

Various 5'O-N-protected deoxynucleoside-3'-O-beta-cyanoethyl-N,N-dialkylamino-/N- morpholinophosphoramidites were prepared from beta-cyanoethyl monochlorophosphoramidites of N,N-dimethylamine, N,N-diisopropylamine and N-morpholine. These active deoxynucleoside phosphates have successfully been used for oligodeoxynucleotide synthesis on controlled pore glass as polymer support and are very suitable for automated DNA-synthesis due to their stability in solution. The intermediate dichloro-beta- cyanoethoxyphosphine can easily be prepared free from any PC1(3) contamination. The active monomers obtained from beta-cyanoethyl monochloro N,N- diisopropylaminophosphoramidites are favoured. Cleavage of the oligonucleotide chain from the polymer support, N-deacylation and deprotection of beta-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.

Complete cDNA sequence of a Dictyostelium ubiquitin with a carboxy‐terminal tail and identification of the protein using an anti‐peptide antibody

“…The inserts were subcloned into vectors Ml3mp18/19 or pUCl9 and sequenced according to Sanger et al [8] using uni-primer or l&mer oligonucleotide primers as indicated by blocks in fig.2. Oligonucleotides for sequencing and Northern blotting were synthesized by the phosphoamidite method [9] on an Applied Biosystems DNA synthesizer and purified by HPLC.…”

Sequence specificity of the non-natural pyrido[2,3-d]pyrimidine nucleoside in triple helix formation

“…Synthesis of oligodeoxyribonucleotides Oligodeoxyribonucleotides were synthesized on an Applied Biosystems Model 380B DNA synthesizer using standard solidphase ,B-cyanoethyl phosphoramidite chemistry (26,27 on an AST Premium 386/33 computer was used to integrate the volume of each rectangle.…”

Chicken Vigilin Gene organization and expression pattern

“…First, domains which are encoded by a single exon (exons 5 -7 and 16 -18), second, domains which are encoded by a set of two exons; these exons are separated by a randomly inserted intron, which is, nevertheless, always within DNA stretches which code for subdomain A (exons [8][9][10][11][12][13][14][15][19][20][21][22][23], and third, domains 13 and 14, portions of which are found in the most probably fused exon 24 (Fig. 2 B).…”

“…The experimental protocols strictly followed the instructions of the manufacturers. Sequence-specific oligonucleotides used as internal sequencing primers were synthesized on a MilliGen/Biosearch 7500 DNA synthesizer [13].…”