End attachment of phenol-oligonucleotide conjugates to diazotized cellulose

The synthesis of a novel phosphoramidite reagent with a hexanediol backbone is described. This reagent has been used to incorporate a phenol moiety on an oligonucleotide (ODN) directly in the course of its automated synthesis. Multiple phenol attachments can be achieved by repetitive coupling cycles...

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Bibliographic Details
Published in:Bioconjugate chemistry 1993-09, Vol.4 (5), p.380-385
Main Authors: Fontanel, Marie Laurence, Bazin, Herve, Teoule, Robert
Format: Article
Language:English
Subjects:
Analytical, structural and metabolic biochemistry
Base Sequence
Biological and medical sciences
Cellulose - chemistry
Diazonium Compounds - chemistry
Fundamental and applied biological sciences. Psychology
General aspects, investigation methods
Indicators and Reagents
Iodine Radioisotopes
Molecular Sequence Data
Nucleic acids
Oligonucleotides - chemistry
Phenols - chemistry
Phosphorus Radioisotopes
Polynucleotide 5'-Hydroxyl-Kinase - metabolism
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Description
Summary:The synthesis of a novel phosphoramidite reagent with a hexanediol backbone is described. This reagent has been used to incorporate a phenol moiety on an oligonucleotide (ODN) directly in the course of its automated synthesis. Multiple phenol attachments can be achieved by repetitive coupling cycles. A simple and rapid immobilization method is described where phenol-modified ODNs are covalently attached to diazotized cellulose. The binding capacity of the membrane can be modulated, depending on the ODN concentration used, to ca. 180 pmol/cm2. There is at least 80% end attachment of the ODN through the phenol group. In addition, the phenol residue can be used as a carrier for the radiolabeling with 125I. The non-nucleosidic hexanediol derivative incorporated at the 5'-end of the ODN is recognized as a substrate by the T4 polynucleotide kinase and the terminal hydroxyl group is successfully phosphorylated allowing its 32P labeling.
ISSN:1043-1802
1520-4812
DOI:10.1021/bc00023a013