Simplification of DNA synthesis by the phosphotriester method

Cashion, P.; Porter, Kenneth Wiggins; Cadger, T.; Sathe, Ganesh M.; Tranquilla, T.; Notman, H.J.; Jay, Ernest

doi:10.1016/s0040-4039(00)93105-8

Cited by 19 publications

(2 citation statements)

References 17 publications

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“…This clearly shows that (3'+3') linked oligonucleotides were either not formed in the synthesis or they were formed in minute quantities such that their presence could not be detected even by the very sensitive procedure described above. Our results are, in fact, in agreement with the observations of Cashion et al 12 and in disagreement with the observations of Letsinger et al3. Although we do not have a definite explanation for such discrepancy, we believe that the p-chlorophenyl group in some fashion provides selectivity in (5'+3') internucleotide bond formation.…”

Section: Resultssupporting

confidence: 79%

Chemical synthesis of a self-complementary octanucleotide, dG-G-T-T-A-A-C-C by a modified triester method

Agarwal¹,

Riftina²

1978

Nucl Acids Res

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The synthesis of a self-complementary octanucleotide, d(G-G-T-T-A-A-C-C-), using a modified triester approach is described. The protected dinucleotides, d(Me2O)TribG(C1C6H4) ibG, d(Me2O)TrT(ClC6H4)T, d(Me2O)TrbzA(ClC6H4)bzA, and d(Me2O)TranC(ClC6H4)anC were synthesized by a one step triester procedure. After removal of the trityl group, the dinucleotides, dT(ClC6H4)T and danC (ClC6H4)anC were coupled to d(Me2O)TribG(ClC6H4)ibG and d(Me2O)TrbzA(ClC6H4)bzA, respectively to yield the respective tetranucleotides. The tetranucleotide, d(Me2O)TrbzA(ClC6H4)bzA(ClC6H4) and (ClC6H4)anC was detritylated and then coupled with d(Me2O)TribG(ClC6H4)ibG(ClC6H4)T(Cl6H4)T to give octanucleotide. The fully protected octanucleotide was deblocked by treatment with aqueous NH3 followed by acid and was characterized by nucleotide sequence analysis.

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Section: Resultssupporting

confidence: 79%

Chemical synthesis of a self-complementary octanucleotide, dG-G-T-T-A-A-C-C by a modified triester method

Agarwal¹,

Riftina²

1978

Nucl Acids Res

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“…Most homoand hetero-polynucleotodes were from Sigma Chemical Co., otherwise, they were synthesized by us with PNP'ase as described below. All synthetic deoxypolymers were made according to established triester methods (5,12); 3H-FmRNA (5,000 CPM/ig) and 3H-rRNA (2980 CPM/ig) isolated from B. mori (13) were supplied by Dr. P. Lizardi. dT cellulose (T3) was from Collaborative Research, Waltham, Mass.…”

Section: Methodsmentioning

confidence: 99%

Hydrophobic affinity chromatography of nucleic acids and proteins

et al. 1980

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5' tritylated oligonucleotides binding hydrophobically to low trityl cellulose/sepharose (< 15 microMTr/ml) retain their hydrogen-bonding specificities for complementary sequences. This, constitutes a novel mode of attaching affinity ligands to solid supports, is more convenient than existing methods, and proceeds with 100% yield. The salt, dielectric constant and temperature dependence of these non-covalently anchored ligands permits the isolation of a variety of RNAs including fibroin mRNA. Medium trityl sepharose (15-40 microM Tr/ml) has a high binding specificity for poly A and poly A containing mRNA, equivalent to dT cellulose. Most proteins, including nucleic acid enzymes, bind to these columns and retain enzymatic activity, thus mimicking enzymes attached covalently to solid phases. A number of in vivo counterparts to this hydrophobically determined specificity are noted, as are homologies to nitro-cellulose filters.

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