Rapid synthesis of long-chain deoxyribooligonucleotides by the N-methylimidazolide phosphotriester method
نویسندگان
چکیده
منابع مشابه
Oligonucleotide synthesis using the manual phosphotriester method.
During the past few years, synthetic DNA, used as a primer in DNA polymerization, in site-directed mutagenesis or as a probe in gene selection, has assumed a central role in recombinant DNA technology (1). This was made possible by the development of methods for efficient solid phase chemical synthesis. Deoxyribonucleotides are ideally suited to solid-phase synthesis since their relative chemic...
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The synthesis of oligothymidilic acids, (dT)m (where m = 4, 7, 10, 13, 16, 19, 22, and 25), was carried out using a solid phase approach in combination with the modified phosphotriester methodology developed in solution. Cellulose was used as the solid support after its functionalization with a specially featured dinucleoside diphosphate, 5'-0-p-chlorophenylphospho-2'(3')-0-acetyluridilyl-[2'(3...
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A new polymer supported synthesis of deoxyribooligonucleotides which can be adapted to automation is described. The method is based on the elongation of an oligomer chain in the 5'- to 3' -end direction using the modified phosphotriester chemistry. The approach is exemplified by the synthesis of a nonanucleotide d(TTCGTCTTG).
متن کاملFurther improvements on the phosphotriester synthesis of deoxyribooligonucleotides and the oligonucleotide directed site-specific mutagenesis of E. coli lipoprotein gene.
Two improvements that greatly enhance the rate of phosphotriester oligonucleotide synthesis are described: 1) use of hindered primary amines, e.g. t-butyl amine for decyanoethylation of oligonucleotide triester intermediates, and 2) a simplified isolation procedure that eliminates the tedious bicarbonate extraction after each condensing reaction. Using the improved procedures, oligonucleotide f...
متن کاملSynthesis of short RNA fragments by the benzotriazolyl phosphotriester approach.
Analytical and experimental evidence will be presented to show that addition of a slight excess of 2-chlorophenyl-0,0-bis[1-benzotriazolyl]phosphate to a properly 2',5'-protected ribonucleoside gives an intermediate 3'-phosphotriester, which, after addition of a 2'-protected ribonucleoside, affords a dimer containing solely a 3'-5'-internucleotide linkage. The above phosphorylation procedure ha...
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ژورنال
عنوان ژورنال: Nucleic Acids Research
سال: 1983
ISSN: 0305-1048,1362-4962
DOI: 10.1093/nar/11.23.8369