Skip to main content
NCBI home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1988 Nov 25;16(22):10861–10880. doi: 10.1093/nar/16.22.10861

Assessment of methods for covalent binding of nucleic acids to magnetic beads, Dynabeads, and the characteristics of the bound nucleic acids in hybridization reactions.

V Lund 1, R Schmid 1, D Rickwood 1, E Hornes 1
PMCID: PMC338944  PMID: 3205723

Abstract

Dynabeads are magnetic monosized beads with high stability, high uniformity, unique paramagnetic properties, low particle-particle interaction, and high dispersibility. Different reactive groups; hydroxyl, carboxyl and amino groups can be attached to the surface. Several methods for covalent attachment of DNA or oligonucleotides to the beads were investigated. Best coupling yields were obtained by carbodiimide-mediated end-attachment of 5'-phosphate and 5'-NH2 modified nucleic acids to respectively amino and carboxyl beads. The carboxyl beads showed a low degree of non-specific binding, while a better yield of end-attached nucleic acids was obtained using the amino beads. The DNA-beads worked efficiently in hybridization experiments, and the kinetics of hybridization approach those of solution hybridization.

Full text

PDF
10861

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Agrawal S., Christodoulou C., Gait M. J. Efficient methods for attaching non-radioactive labels to the 5' ends of synthetic oligodeoxyribonucleotides. Nucleic Acids Res. 1986 Aug 11;14(15):6227–6245. doi: 10.1093/nar/14.15.6227. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Arndt-Jovin D. J., Jovin T. M., Bähr W., Frischauf A. M., Marquardt M. Covalent attachment of DNA to agarose. Improved synthesis and use in affinity chromatography. Eur J Biochem. 1975 Jun;54(2):411–418. doi: 10.1111/j.1432-1033.1975.tb04151.x. [DOI] [PubMed] [Google Scholar]
  3. Bünemann H., Westhoff P., Herrmann R. G. Immobilization of denatured DNA to macroporous supports: I. Efficiency of different coupling procedures. Nucleic Acids Res. 1982 Nov 25;10(22):7163–7180. doi: 10.1093/nar/10.22.7163. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Chu B. C., Orgel L. E. Detection of specific DNA sequences with short biotin-labeled probes. DNA. 1985 Aug;4(4):327–331. doi: 10.1089/dna.1985.4.327. [DOI] [PubMed] [Google Scholar]
  5. Chu B. C., Orgel L. E. Nonenzymatic sequence-specific cleavage of single-stranded DNA. Proc Natl Acad Sci U S A. 1985 Feb;82(4):963–967. doi: 10.1073/pnas.82.4.963. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Chu B. C., Wahl G. M., Orgel L. E. Derivatization of unprotected polynucleotides. Nucleic Acids Res. 1983 Sep 24;11(18):6513–6529. doi: 10.1093/nar/11.18.6513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Clerici L., Campagnari F., de Rooij J. F., van Boom J. H. Preparation of polydeoxynucleotides linked to a solid support by coupling CNBr-activated cellulose with 5'-NH2-terminated oligo and poly(pdT)'s. Nucleic Acids Res. 1979 Jan;6(1):247–258. doi: 10.1093/nar/6.1.247. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Connolly B. A. The synthesis of oligonucleotides containing a primary amino group at the 5'-terminus. Nucleic Acids Res. 1987 Apr 10;15(7):3131–3139. doi: 10.1093/nar/15.7.3131. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Ghosh S. S., Musso G. F. Covalent attachment of oligonucleotides to solid supports. Nucleic Acids Res. 1987 Jul 10;15(13):5353–5372. doi: 10.1093/nar/15.13.5353. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gilham P. T. The synthesis of celluloses containing covalently bound nucleotides, polynucleotides, and nucleic acids. Biochemistry. 1968 Aug;7(8):2809–2813. doi: 10.1021/bi00848a016. [DOI] [PubMed] [Google Scholar]
  11. Gingeras T. R., Kwoh D. Y., Davis G. R. Hybridization properties of immobilized nucleic acids. Nucleic Acids Res. 1987 Jul 10;15(13):5373–5390. doi: 10.1093/nar/15.13.5373. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kremsky J. N., Wooters J. L., Dougherty J. P., Meyers R. E., Collins M., Brown E. L. Immobilization of DNA via oligonucleotides containing an aldehyde or carboxylic acid group at the 5' terminus. Nucleic Acids Res. 1987 Apr 10;15(7):2891–2909. doi: 10.1093/nar/15.7.2891. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Moss L. G., Moore J. P., Chan L. A simple, efficient method for coupling DNA to cellulose. Development of the method and application to mRNA purification. J Biol Chem. 1981 Dec 25;256(24):12655–12658. [PubMed] [Google Scholar]
  14. Noyes B. E., Stark G. R. Nucleic acid hybridization using DNA covalently coupled to cellulose. Cell. 1975 Jul;5(3):301–310. doi: 10.1016/0092-8674(75)90105-1. [DOI] [PubMed] [Google Scholar]
  15. Rickwood D. An improved method for the insolubilization of DNA for affinity chromatography. Biochim Biophys Acta. 1972 Apr 26;269(1):47–50. doi: 10.1016/0005-2787(72)90072-x. [DOI] [PubMed] [Google Scholar]
  16. Wolf S. F., Haines L., Fisch J., Kremsky J. N., Dougherty J. P., Jacobs K. Rapid hybridization kinetics of DNA attached to submicron latex particles. Nucleic Acids Res. 1987 Apr 10;15(7):2911–2926. doi: 10.1093/nar/15.7.2911. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES