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. 1997 Dec 1;25(23):4835–4841. doi: 10.1093/nar/25.23.4835

An improved divergent synthesis of comb-type branched oligodeoxyribonucleotides (bDNA) containing multiple secondary sequences.

T Horn 1, C A Chang 1, M S Urdea 1
PMCID: PMC147104  PMID: 9365265

Abstract

The divergent synthesis of branched DNA (bDNA) comb structures is described. This new type of bDNA contains one unique oligonucleotide, the primary sequence, covalently attached through a comb-like branch network to many identical copies of a different oligonucleotide, the secondary sequence. The bDNA comb structures were assembled on a solid support and several synthesis parameters were investigated and optimized. The bDNA comb molecules were characterized by polyacrylamide gel electrophoretic methods and by controlled cleavage at periodate-cleavable moieties incorporated during synthesis. The developed chemistry allows synthesis of bDNA comb molecules containing multiple secondary sequences. In the accompanying article we describe the synthesis and characterization of large bDNA combs containing all four deoxynucleotides for use as signal amplifiers in nucleic acid quantification assays.

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Selected References

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  1. Barany F. Genetic disease detection and DNA amplification using cloned thermostable ligase. Proc Natl Acad Sci U S A. 1991 Jan 1;88(1):189–193. doi: 10.1073/pnas.88.1.189. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Braich R. S., Damha M. J. Regiospecific solid-phase synthesis of branched oligonucleotides. Effect of vicinal 2',5'- (or 2',3'-) and 3',5'-phosphodiester linkages on the formation of hairpin DNA. Bioconjug Chem. 1997 May-Jun;8(3):370–377. doi: 10.1021/bc9700300. [DOI] [PubMed] [Google Scholar]
  3. Horn T., Urdea M. S. Forks and combs and DNA: the synthesis of branched oligodeoxyribonucleotides. Nucleic Acids Res. 1989 Sep 12;17(17):6959–6967. doi: 10.1093/nar/17.17.6959. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Keith G., Gilham P. T. Stepwise degradation of polyribonucleotides. Biochemistry. 1974 Aug 13;13(17):3601–3606. doi: 10.1021/bi00714a031. [DOI] [PubMed] [Google Scholar]
  5. Letsinger R. L., Caruthers M. H., Miller P. S., Ogilvie K. K. Oligonucleotide syntheses utilizing beta-benzoylpropionyl, a blocking group with a trigger for selective cleavage. J Am Chem Soc. 1967 Dec 20;89(26):7146–7147. doi: 10.1021/ja01002a074. [DOI] [PubMed] [Google Scholar]
  6. Letsinger R. L., Miller P. S., Grams G. W. Selective N-debenzoylation of N,O-polybenzoylucleosides. Tetrahedron Lett. 1968 Apr;(22):2621–2624. doi: 10.1016/s0040-4039(00)89658-6. [DOI] [PubMed] [Google Scholar]
  7. Mullis K. B., Faloona F. A. Specific synthesis of DNA in vitro via a polymerase-catalyzed chain reaction. Methods Enzymol. 1987;155:335–350. doi: 10.1016/0076-6879(87)55023-6. [DOI] [PubMed] [Google Scholar]
  8. Persing D. H., Landry M. L. In vitro amplification techniques for the detection of nucleic acids: new tools for the diagnostic laboratory. Yale J Biol Med. 1989 Mar-Apr;62(2):159–171. [PMC free article] [PubMed] [Google Scholar]
  9. Pon R. T., Usman N., Ogilvie K. K. Derivatization of controlled pore glass beads for solid phase oligonucleotide synthesis. Biotechniques. 1988 Sep;6(8):768–775. [PubMed] [Google Scholar]
  10. Urdea M. S., Warner B. D., Running J. A., Stempien M., Clyne J., Horn T. A comparison of non-radioisotopic hybridization assay methods using fluorescent, chemiluminescent and enzyme labeled synthetic oligodeoxyribonucleotide probes. Nucleic Acids Res. 1988 Jun 10;16(11):4937–4956. doi: 10.1093/nar/16.11.4937. [DOI] [PMC free article] [PubMed] [Google Scholar]

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