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Nucleic Acids Research, 1988, Vol. 16, No. 22 10861-10880
© 1988

CHEMISTRY

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

Vera Lund1,4, Ruth Schmid3, David Rickwood4, Federico Robbiati4 and Erik Homes2

1Apothekemes Laboratorium A.S. Tromsø 2Apothekemes Laboratorium A.S. Oslo 3SINTEF, Division of Applied Chemistry, Trondheim, Norway 4Department of Biology, University of Essex UK

Received July 12, 1988. Revised September 28, 1988. Accepted October 21, 1988.

DynabeadsTM 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.


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