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Nucleic Acids Research, 1987, Vol. 15, No. 13 5373-5390
© 1987

Articles

Hybridization properties of immobilized nucleic acids

T.R. Gingeras*, D.Y. Kwoh and G.R. Davis

SISKA Diagnostics, Inc. and The Salk Institute Biotechnology/Industrial Associates Inc., La Jolla, CA 92037, USA

*To whom correspondence should be addressed

Received February 11, 1987. Revised June 2, 1987. Accepted June 2, 1987.

The 5'-end attachment of oligonucleotides to dextran supports facilitates the study of the hybridization properties of an immobilized oligonucleotide system. The hybridization properties which were studied include: hybridization capacity and kinetics, hybridization-complex stability, and reagents influencing hybridization efficiency. Results of these experiments reveal that the hybridization efficiencies of support-bound oligonucleotides were 75–80% and 40–50% for single-stranded oligonucleotide targets and long double-stranded targets, respectively. These hybridization efficiencies are dependent upon prehybridizing the support-bound oligonucleotides with dextran sulfate. In addition, comparisons of the relative hybridization efficiencies of the support-bound oligonucleotide and nitrocellulose-based systems have been made which indicate a retention of 13–28% of target sequences on the filters and a detection efficiency of 8–20%.


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